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Increased expression of human T lymphocyte virus type I (HTLV-I) Tax11-19 peptide-human histocompatibility leukocyte antigen A*201 complexes on CD4+ CD25+ T Cells detected by peptide-specific, major histocompatibility complex-restricted antibodies in patients with HTLV-I-associated neurologic disease.

Yamano Y, Cohen CJ, Takenouchi N, Yao K, Tomaru U, Li HC, Reiter Y, Jacobson S - J. Exp. Med. (2004)

Bottom Line: Human T lymphocyte virus type I (HTLV-I)-associated chronic inflammatory neurological disease (HTLV-I-associated myelopathy/tropical spastic paraparesis [HAM/TSP]) is suggested to be an immunopathologically mediated disorder characterized by large numbers of HTLV-I Tax-specific CD8+ T cells.However, until now, it has not been possible to identify and/or quantify these peptide-HLA complexes.These results indicate that the increased detection and visualization of peptide-HLA complexes in HAM/TSP CD4+ CD25+ T cell subsets that are shown to stimulate and expand HTLV-I Tax-specific CD8+ T cells may play an important role in the pathogenesis of HTLV-I-associated neurological disease.

View Article: PubMed Central - PubMed

Affiliation: Viral Immunology Section, Neuroimmunology Branch, National Institutes of Health, National Institute of Neurological Disorders and Strokes, Building 10, Room 5B-16, Bethesda, MD 20892, USA.

ABSTRACT
Human T lymphocyte virus type I (HTLV-I)-associated chronic inflammatory neurological disease (HTLV-I-associated myelopathy/tropical spastic paraparesis [HAM/TSP]) is suggested to be an immunopathologically mediated disorder characterized by large numbers of HTLV-I Tax-specific CD8+ T cells. The frequency of these cells in the peripheral blood and cerebrospinal fluid is proportional to the amount of HTLV-I proviral load and the levels of HTLV-I tax mRNA expression. As the stimulus for these virus-specific T cells are immunodominant peptide-human histocompatibility leukocyte antigen (HLA) complexes expressed on antigen-presenting cells, it was of interest to determine which cells express these complexes and at what frequency. However, until now, it has not been possible to identify and/or quantify these peptide-HLA complexes. Using a recently developed antibody that specifically recognizes Tax11-19 peptide-HLA-A*201 complexes, the level of Tax11-19-HLA-A*201 expression on T cells was demonstrated to be increased in HAM/TSP and correlated with HTLV-I proviral DNA load, HTLV-I tax mRNA load, and HTLV-I Tax-specific CD8+ T cell frequencies. Furthermore, CD4+ CD25+ T cells were demonstrated to be the major reservoir of HTLV-I provirus as well as Tax11-19 peptide-HLA-A*201 complexes. These results indicate that the increased detection and visualization of peptide-HLA complexes in HAM/TSP CD4+ CD25+ T cell subsets that are shown to stimulate and expand HTLV-I Tax-specific CD8+ T cells may play an important role in the pathogenesis of HTLV-I-associated neurological disease.

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Peptide-specific, HLA-restricted binding of Ab. (A) Peptide-specific binding of anti–Tax11-19 peptide–HLA-A*201 Ab (TaxA2-Ab) on HmyA2.1 cells. HmyA2.1 cells were pulsed with HTLV-I Tax11-19 peptide or control melanoma gp100–derived G9-154 peptides, incubated with PE-labeled TaxA2-Ab, and then analyzed by flow cytometry. TaxA2-Ab reacted only with Tax11-19 peptide–loaded HmyA2.1 cells (solid line), but not with cells loaded with the control peptide (dotted line). (B) Peptide-specific binding of anti–melanoma gp100 G9-154 peptide–HLA-A*201 Ab (MelanomaA2-Ab) on HmyA2.1 cells. HmyA2.1 cells were pulsed with melanoma gp100–derived G9-154 peptides or control HTLV-I Tax11-19 peptide, incubated with PE-labeled MelanomaA2-Ab, and then analyzed by flow cytometry. MelanomaA2-Ab reacted only with G9-154 peptide–loaded HmyA2.1 cells (solid line), but not with cells loaded with the control peptide (dotted line). (C) Peptide titration study of TaxA2-Ab staining on HmyA2.1 cells. HmyA2.1 cells were pulsed with the indicated concentration of HTLV-I Tax11-19 peptide, incubated with PE-labeled TaxA2-Ab, and then analyzed by flow cytometry. The levels of Tax11-19 peptide–HLA-A*201 complexes are expressed by mean fluorescence intensity. (D and E) Peptide-specific binding of TaxA2-Ab on CD4+ T cells (D) and CD8+ T cells (E) from an HLA-A*201+ healthy donor (A2HD). CD4+ and CD8+ T cells from A2HD were pulsed with 100 μM HTLV-I Tax11-19 peptide or control melanoma gp100–derived G9-154 peptides, incubated with PE-labeled TaxA2-Ab, and then analyzed by flow cytometry. TaxA2-Ab reacted only with Tax11-19 peptide–loaded HmyA2.1 cells (solid line), but not with cells loaded with the control peptide (dotted line). (F) Peptide titration study of TaxA2-Ab staining on CD4+ and CD8+ T cells from A2HD. CD4+ (•) and CD8+ (▪) T cells from A2HD were pulsed with the indicated concentration of HTLV-I Tax11-19 peptide, incubated with PE-labeled TaxA2-Ab, and then analyzed by flow cytometry. The levels of Tax11-19 peptide–HLA-A*201 complexes are expressed by mean fluorescence intensity.
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fig1: Peptide-specific, HLA-restricted binding of Ab. (A) Peptide-specific binding of anti–Tax11-19 peptide–HLA-A*201 Ab (TaxA2-Ab) on HmyA2.1 cells. HmyA2.1 cells were pulsed with HTLV-I Tax11-19 peptide or control melanoma gp100–derived G9-154 peptides, incubated with PE-labeled TaxA2-Ab, and then analyzed by flow cytometry. TaxA2-Ab reacted only with Tax11-19 peptide–loaded HmyA2.1 cells (solid line), but not with cells loaded with the control peptide (dotted line). (B) Peptide-specific binding of anti–melanoma gp100 G9-154 peptide–HLA-A*201 Ab (MelanomaA2-Ab) on HmyA2.1 cells. HmyA2.1 cells were pulsed with melanoma gp100–derived G9-154 peptides or control HTLV-I Tax11-19 peptide, incubated with PE-labeled MelanomaA2-Ab, and then analyzed by flow cytometry. MelanomaA2-Ab reacted only with G9-154 peptide–loaded HmyA2.1 cells (solid line), but not with cells loaded with the control peptide (dotted line). (C) Peptide titration study of TaxA2-Ab staining on HmyA2.1 cells. HmyA2.1 cells were pulsed with the indicated concentration of HTLV-I Tax11-19 peptide, incubated with PE-labeled TaxA2-Ab, and then analyzed by flow cytometry. The levels of Tax11-19 peptide–HLA-A*201 complexes are expressed by mean fluorescence intensity. (D and E) Peptide-specific binding of TaxA2-Ab on CD4+ T cells (D) and CD8+ T cells (E) from an HLA-A*201+ healthy donor (A2HD). CD4+ and CD8+ T cells from A2HD were pulsed with 100 μM HTLV-I Tax11-19 peptide or control melanoma gp100–derived G9-154 peptides, incubated with PE-labeled TaxA2-Ab, and then analyzed by flow cytometry. TaxA2-Ab reacted only with Tax11-19 peptide–loaded HmyA2.1 cells (solid line), but not with cells loaded with the control peptide (dotted line). (F) Peptide titration study of TaxA2-Ab staining on CD4+ and CD8+ T cells from A2HD. CD4+ (•) and CD8+ (▪) T cells from A2HD were pulsed with the indicated concentration of HTLV-I Tax11-19 peptide, incubated with PE-labeled TaxA2-Ab, and then analyzed by flow cytometry. The levels of Tax11-19 peptide–HLA-A*201 complexes are expressed by mean fluorescence intensity.

Mentions: To demonstrate the specificity of peptide-specific, MHC-restricted Fab Abs that bind to Tax11-19 peptide–HLA-A*201 complexes (TaxA2-Ab), a human immortalized B cell line expressing HLA-A*201 (HmyA2.1) was pulsed with 10 μM HTLV-I Tax11-19 peptide or control melanoma gp100–derived G9-154 peptides, and incubated with PE-labeled TaxA2-Ab. The ability of TaxA2-Ab to bind to Tax11-19 peptide–HLA-A*201 molecule was then monitored by flow cytometry. As shown in Fig. 1 A, TaxA2-Ab reacted only with Tax11-19 peptide–loaded HmyA2.1 cells, but not with cells loaded with the control peptide. Conversely, a peptide-specific, MHC-restricted Fab Ab specific for the melanoma gp100 G9-154 peptide–HLA-A*201 complex (MelanomaA2-Ab) specifically bound HmyA2.1 cells pulsed with the melanoma gp100 peptide, but not Tax11-19 (Fig. 1 B). Peptide titration studies demonstrated that the level of TaxA2-Ab staining on HmyA2.1 cells correlated with the concentration of loaded Tax11-19 peptide (Fig. 1 C).


Increased expression of human T lymphocyte virus type I (HTLV-I) Tax11-19 peptide-human histocompatibility leukocyte antigen A*201 complexes on CD4+ CD25+ T Cells detected by peptide-specific, major histocompatibility complex-restricted antibodies in patients with HTLV-I-associated neurologic disease.

Yamano Y, Cohen CJ, Takenouchi N, Yao K, Tomaru U, Li HC, Reiter Y, Jacobson S - J. Exp. Med. (2004)

Peptide-specific, HLA-restricted binding of Ab. (A) Peptide-specific binding of anti–Tax11-19 peptide–HLA-A*201 Ab (TaxA2-Ab) on HmyA2.1 cells. HmyA2.1 cells were pulsed with HTLV-I Tax11-19 peptide or control melanoma gp100–derived G9-154 peptides, incubated with PE-labeled TaxA2-Ab, and then analyzed by flow cytometry. TaxA2-Ab reacted only with Tax11-19 peptide–loaded HmyA2.1 cells (solid line), but not with cells loaded with the control peptide (dotted line). (B) Peptide-specific binding of anti–melanoma gp100 G9-154 peptide–HLA-A*201 Ab (MelanomaA2-Ab) on HmyA2.1 cells. HmyA2.1 cells were pulsed with melanoma gp100–derived G9-154 peptides or control HTLV-I Tax11-19 peptide, incubated with PE-labeled MelanomaA2-Ab, and then analyzed by flow cytometry. MelanomaA2-Ab reacted only with G9-154 peptide–loaded HmyA2.1 cells (solid line), but not with cells loaded with the control peptide (dotted line). (C) Peptide titration study of TaxA2-Ab staining on HmyA2.1 cells. HmyA2.1 cells were pulsed with the indicated concentration of HTLV-I Tax11-19 peptide, incubated with PE-labeled TaxA2-Ab, and then analyzed by flow cytometry. The levels of Tax11-19 peptide–HLA-A*201 complexes are expressed by mean fluorescence intensity. (D and E) Peptide-specific binding of TaxA2-Ab on CD4+ T cells (D) and CD8+ T cells (E) from an HLA-A*201+ healthy donor (A2HD). CD4+ and CD8+ T cells from A2HD were pulsed with 100 μM HTLV-I Tax11-19 peptide or control melanoma gp100–derived G9-154 peptides, incubated with PE-labeled TaxA2-Ab, and then analyzed by flow cytometry. TaxA2-Ab reacted only with Tax11-19 peptide–loaded HmyA2.1 cells (solid line), but not with cells loaded with the control peptide (dotted line). (F) Peptide titration study of TaxA2-Ab staining on CD4+ and CD8+ T cells from A2HD. CD4+ (•) and CD8+ (▪) T cells from A2HD were pulsed with the indicated concentration of HTLV-I Tax11-19 peptide, incubated with PE-labeled TaxA2-Ab, and then analyzed by flow cytometry. The levels of Tax11-19 peptide–HLA-A*201 complexes are expressed by mean fluorescence intensity.
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Related In: Results  -  Collection

Show All Figures
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fig1: Peptide-specific, HLA-restricted binding of Ab. (A) Peptide-specific binding of anti–Tax11-19 peptide–HLA-A*201 Ab (TaxA2-Ab) on HmyA2.1 cells. HmyA2.1 cells were pulsed with HTLV-I Tax11-19 peptide or control melanoma gp100–derived G9-154 peptides, incubated with PE-labeled TaxA2-Ab, and then analyzed by flow cytometry. TaxA2-Ab reacted only with Tax11-19 peptide–loaded HmyA2.1 cells (solid line), but not with cells loaded with the control peptide (dotted line). (B) Peptide-specific binding of anti–melanoma gp100 G9-154 peptide–HLA-A*201 Ab (MelanomaA2-Ab) on HmyA2.1 cells. HmyA2.1 cells were pulsed with melanoma gp100–derived G9-154 peptides or control HTLV-I Tax11-19 peptide, incubated with PE-labeled MelanomaA2-Ab, and then analyzed by flow cytometry. MelanomaA2-Ab reacted only with G9-154 peptide–loaded HmyA2.1 cells (solid line), but not with cells loaded with the control peptide (dotted line). (C) Peptide titration study of TaxA2-Ab staining on HmyA2.1 cells. HmyA2.1 cells were pulsed with the indicated concentration of HTLV-I Tax11-19 peptide, incubated with PE-labeled TaxA2-Ab, and then analyzed by flow cytometry. The levels of Tax11-19 peptide–HLA-A*201 complexes are expressed by mean fluorescence intensity. (D and E) Peptide-specific binding of TaxA2-Ab on CD4+ T cells (D) and CD8+ T cells (E) from an HLA-A*201+ healthy donor (A2HD). CD4+ and CD8+ T cells from A2HD were pulsed with 100 μM HTLV-I Tax11-19 peptide or control melanoma gp100–derived G9-154 peptides, incubated with PE-labeled TaxA2-Ab, and then analyzed by flow cytometry. TaxA2-Ab reacted only with Tax11-19 peptide–loaded HmyA2.1 cells (solid line), but not with cells loaded with the control peptide (dotted line). (F) Peptide titration study of TaxA2-Ab staining on CD4+ and CD8+ T cells from A2HD. CD4+ (•) and CD8+ (▪) T cells from A2HD were pulsed with the indicated concentration of HTLV-I Tax11-19 peptide, incubated with PE-labeled TaxA2-Ab, and then analyzed by flow cytometry. The levels of Tax11-19 peptide–HLA-A*201 complexes are expressed by mean fluorescence intensity.
Mentions: To demonstrate the specificity of peptide-specific, MHC-restricted Fab Abs that bind to Tax11-19 peptide–HLA-A*201 complexes (TaxA2-Ab), a human immortalized B cell line expressing HLA-A*201 (HmyA2.1) was pulsed with 10 μM HTLV-I Tax11-19 peptide or control melanoma gp100–derived G9-154 peptides, and incubated with PE-labeled TaxA2-Ab. The ability of TaxA2-Ab to bind to Tax11-19 peptide–HLA-A*201 molecule was then monitored by flow cytometry. As shown in Fig. 1 A, TaxA2-Ab reacted only with Tax11-19 peptide–loaded HmyA2.1 cells, but not with cells loaded with the control peptide. Conversely, a peptide-specific, MHC-restricted Fab Ab specific for the melanoma gp100 G9-154 peptide–HLA-A*201 complex (MelanomaA2-Ab) specifically bound HmyA2.1 cells pulsed with the melanoma gp100 peptide, but not Tax11-19 (Fig. 1 B). Peptide titration studies demonstrated that the level of TaxA2-Ab staining on HmyA2.1 cells correlated with the concentration of loaded Tax11-19 peptide (Fig. 1 C).

Bottom Line: Human T lymphocyte virus type I (HTLV-I)-associated chronic inflammatory neurological disease (HTLV-I-associated myelopathy/tropical spastic paraparesis [HAM/TSP]) is suggested to be an immunopathologically mediated disorder characterized by large numbers of HTLV-I Tax-specific CD8+ T cells.However, until now, it has not been possible to identify and/or quantify these peptide-HLA complexes.These results indicate that the increased detection and visualization of peptide-HLA complexes in HAM/TSP CD4+ CD25+ T cell subsets that are shown to stimulate and expand HTLV-I Tax-specific CD8+ T cells may play an important role in the pathogenesis of HTLV-I-associated neurological disease.

View Article: PubMed Central - PubMed

Affiliation: Viral Immunology Section, Neuroimmunology Branch, National Institutes of Health, National Institute of Neurological Disorders and Strokes, Building 10, Room 5B-16, Bethesda, MD 20892, USA.

ABSTRACT
Human T lymphocyte virus type I (HTLV-I)-associated chronic inflammatory neurological disease (HTLV-I-associated myelopathy/tropical spastic paraparesis [HAM/TSP]) is suggested to be an immunopathologically mediated disorder characterized by large numbers of HTLV-I Tax-specific CD8+ T cells. The frequency of these cells in the peripheral blood and cerebrospinal fluid is proportional to the amount of HTLV-I proviral load and the levels of HTLV-I tax mRNA expression. As the stimulus for these virus-specific T cells are immunodominant peptide-human histocompatibility leukocyte antigen (HLA) complexes expressed on antigen-presenting cells, it was of interest to determine which cells express these complexes and at what frequency. However, until now, it has not been possible to identify and/or quantify these peptide-HLA complexes. Using a recently developed antibody that specifically recognizes Tax11-19 peptide-HLA-A*201 complexes, the level of Tax11-19-HLA-A*201 expression on T cells was demonstrated to be increased in HAM/TSP and correlated with HTLV-I proviral DNA load, HTLV-I tax mRNA load, and HTLV-I Tax-specific CD8+ T cell frequencies. Furthermore, CD4+ CD25+ T cells were demonstrated to be the major reservoir of HTLV-I provirus as well as Tax11-19 peptide-HLA-A*201 complexes. These results indicate that the increased detection and visualization of peptide-HLA complexes in HAM/TSP CD4+ CD25+ T cell subsets that are shown to stimulate and expand HTLV-I Tax-specific CD8+ T cells may play an important role in the pathogenesis of HTLV-I-associated neurological disease.

Show MeSH
Related in: MedlinePlus