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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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Increased detection of Tax11-19 peptide–HLA-A*201 complexes on CD4+ T cells in HAM/TSP patients. (A) Representative histograms of increased sensitivity of CD4+ T cells from HAM/TSP patients for TaxA2-Ab staining. Fluorescence intensity for TaxA2-Ab staining of CD4+ T cells from a HAM/TSP patient (bottom left) was stronger than CD4+ T cells from A2HD (top left) when pulsed with 10 μM Tax11-19 peptide (solid line) compared with 10 μM control melanoma gp100 G9-154 peptide (dotted line). However, fluorescence intensity for MelanomaA2-Ab staining of CD4+ T cells from a HAM/TSP patient (bottom right) was similar to that of A2HD CD4+ T cells (top right) when pulsed with 10 μM melanoma gp100 G9-154 peptide (solid line) compared with 10 μM control Tax11-19 peptide (dotted line). A2HD, HLA-A*201+ healthy donor. (B) Fluorescence intensity for TaxA2-Ab staining of CD4+ T cells from six HAM/TSP patients was significantly stronger (P = 0.0039) than CD4+ T cells from six A2HD patients when pulsed with 10 μM Tax11-19 peptide. (C) No significant difference (P = 0.1093) of the fluorescence intensity of anti–HLA-A*201 staining between CD4+ T cells from six HAM/TSP patients and CD4+ T cells from six A2HD patients. (D) Correlation between the level of Tax11-19 peptide–HLA-A*201 expression and HTLV-I DNA load, RNA load, and HTLV-I Tax–specific CD8+ T cell frequency. The levels of Tax11-19 peptide–HLA-A*201 expression on CD4+ T cells were significantly correlated with HTLV-I proviral DNA load (P = 0.0490, r2 = 0.661), HTLV-I mRNA load (P = 0.0259, r2 = 0.749), and HTLV-I Tax11-19 tetramer–specific CD8+ T cell frequencies (P = 0.0470, r2 = 0.668) in six HAM/TSP patients.
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fig5: Increased detection of Tax11-19 peptide–HLA-A*201 complexes on CD4+ T cells in HAM/TSP patients. (A) Representative histograms of increased sensitivity of CD4+ T cells from HAM/TSP patients for TaxA2-Ab staining. Fluorescence intensity for TaxA2-Ab staining of CD4+ T cells from a HAM/TSP patient (bottom left) was stronger than CD4+ T cells from A2HD (top left) when pulsed with 10 μM Tax11-19 peptide (solid line) compared with 10 μM control melanoma gp100 G9-154 peptide (dotted line). However, fluorescence intensity for MelanomaA2-Ab staining of CD4+ T cells from a HAM/TSP patient (bottom right) was similar to that of A2HD CD4+ T cells (top right) when pulsed with 10 μM melanoma gp100 G9-154 peptide (solid line) compared with 10 μM control Tax11-19 peptide (dotted line). A2HD, HLA-A*201+ healthy donor. (B) Fluorescence intensity for TaxA2-Ab staining of CD4+ T cells from six HAM/TSP patients was significantly stronger (P = 0.0039) than CD4+ T cells from six A2HD patients when pulsed with 10 μM Tax11-19 peptide. (C) No significant difference (P = 0.1093) of the fluorescence intensity of anti–HLA-A*201 staining between CD4+ T cells from six HAM/TSP patients and CD4+ T cells from six A2HD patients. (D) Correlation between the level of Tax11-19 peptide–HLA-A*201 expression and HTLV-I DNA load, RNA load, and HTLV-I Tax–specific CD8+ T cell frequency. The levels of Tax11-19 peptide–HLA-A*201 expression on CD4+ T cells were significantly correlated with HTLV-I proviral DNA load (P = 0.0490, r2 = 0.661), HTLV-I mRNA load (P = 0.0259, r2 = 0.749), and HTLV-I Tax11-19 tetramer–specific CD8+ T cell frequencies (P = 0.0470, r2 = 0.668) in six HAM/TSP patients.

Mentions: The inability to detect Tax11-19 peptide–HLA-A*201 complexes directly from ex vivo T cells of HAM/TSP patients (Fig. 2 A) could be reflective of a low concentration of Tax11-19 peptide bound to endogenous HLA-A*201 molecules. As we had shown that the staining intensity of the TaxA2-Ab was dependent on the concentration of the peptide used for pulsing T cells in A2HD (Fig. 1 F), it was of interest to determine whether this TaxA2-Ab staining of peptide-pulsed ex vivo T cells was different between A2HD and HLA-A*201 HAM/TSP patients. CD4+ T cells were separated from ex vivo peripheral blood T cells of six A2HD and six HLA-A*201 HAM/TSP patients. These ex vivo T cells were pulsed at a concentration of 10 μM Tax11-19 peptide, previously determined to be in a range where the TaxA2-Ab was unable to detect Tax11-19 peptide–HLA-A*201 complexes (Fig. 1 F). In contrast to HTLV-I Tax11-19 peptide–pulsed A2HD T cells (Figs. 5 A and 1 F), staining intensity of the TaxA2-Ab on comparably peptide-pulsed HAM/TSP T cells was significantly higher (Fig. 5, A and B). To determine if this difference between A2HD and HAM/TSP patients was specific for the amount of Tax11-19 peptide–HLA-A*201 complexes as defined by the TaxA2-Ab, CD4+ T cells from the same individuals were pulsed with the same concentration of control melanoma gp100 G9-154 peptide, and the level of G9-154 peptide–HLA-A*201 complexes was analyzed using the MelanomaA2-Ab. As shown in Fig. 5 A, staining intensity with the MelanomaA2-Ab in HAM/TSP patients was similar to that in A2HD. These observations were confirmed using a wider range of peptide concentrations (Fig. 6, A and B). As the staining sensitivity of the TaxA2-Ab might be affected by the amount of expressed HLA-A*201 complexes, we compared the amount of HLA-A*201 expression on CD4+ T cells between A2HD and HAM/TSP using anti–HLA-A*201–specific Ab (BB7.2). There was no statistically significant difference in the amount of HLA-A*201 expression between A2HD and HAM/TSP (Fig. 5 C), suggesting that differential levels of HLA-A*201 expression do not contribute to the TaxA2-Ab staining intensity between A2HD and HAM/TSP patients. In addition, we were able to identify two HLA-A*0201 asymptomatic carriers from which sufficient cells were available to analyze with the TaxA2-Ab. TaxA2-Ab staining intensities on CD4+ T cells of these two HLA-A*0201 asymptomatic carriers were similar with those on A2HD (unpublished data).


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)

Increased detection of Tax11-19 peptide–HLA-A*201 complexes on CD4+ T cells in HAM/TSP patients. (A) Representative histograms of increased sensitivity of CD4+ T cells from HAM/TSP patients for TaxA2-Ab staining. Fluorescence intensity for TaxA2-Ab staining of CD4+ T cells from a HAM/TSP patient (bottom left) was stronger than CD4+ T cells from A2HD (top left) when pulsed with 10 μM Tax11-19 peptide (solid line) compared with 10 μM control melanoma gp100 G9-154 peptide (dotted line). However, fluorescence intensity for MelanomaA2-Ab staining of CD4+ T cells from a HAM/TSP patient (bottom right) was similar to that of A2HD CD4+ T cells (top right) when pulsed with 10 μM melanoma gp100 G9-154 peptide (solid line) compared with 10 μM control Tax11-19 peptide (dotted line). A2HD, HLA-A*201+ healthy donor. (B) Fluorescence intensity for TaxA2-Ab staining of CD4+ T cells from six HAM/TSP patients was significantly stronger (P = 0.0039) than CD4+ T cells from six A2HD patients when pulsed with 10 μM Tax11-19 peptide. (C) No significant difference (P = 0.1093) of the fluorescence intensity of anti–HLA-A*201 staining between CD4+ T cells from six HAM/TSP patients and CD4+ T cells from six A2HD patients. (D) Correlation between the level of Tax11-19 peptide–HLA-A*201 expression and HTLV-I DNA load, RNA load, and HTLV-I Tax–specific CD8+ T cell frequency. The levels of Tax11-19 peptide–HLA-A*201 expression on CD4+ T cells were significantly correlated with HTLV-I proviral DNA load (P = 0.0490, r2 = 0.661), HTLV-I mRNA load (P = 0.0259, r2 = 0.749), and HTLV-I Tax11-19 tetramer–specific CD8+ T cell frequencies (P = 0.0470, r2 = 0.668) in six HAM/TSP patients.
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fig5: Increased detection of Tax11-19 peptide–HLA-A*201 complexes on CD4+ T cells in HAM/TSP patients. (A) Representative histograms of increased sensitivity of CD4+ T cells from HAM/TSP patients for TaxA2-Ab staining. Fluorescence intensity for TaxA2-Ab staining of CD4+ T cells from a HAM/TSP patient (bottom left) was stronger than CD4+ T cells from A2HD (top left) when pulsed with 10 μM Tax11-19 peptide (solid line) compared with 10 μM control melanoma gp100 G9-154 peptide (dotted line). However, fluorescence intensity for MelanomaA2-Ab staining of CD4+ T cells from a HAM/TSP patient (bottom right) was similar to that of A2HD CD4+ T cells (top right) when pulsed with 10 μM melanoma gp100 G9-154 peptide (solid line) compared with 10 μM control Tax11-19 peptide (dotted line). A2HD, HLA-A*201+ healthy donor. (B) Fluorescence intensity for TaxA2-Ab staining of CD4+ T cells from six HAM/TSP patients was significantly stronger (P = 0.0039) than CD4+ T cells from six A2HD patients when pulsed with 10 μM Tax11-19 peptide. (C) No significant difference (P = 0.1093) of the fluorescence intensity of anti–HLA-A*201 staining between CD4+ T cells from six HAM/TSP patients and CD4+ T cells from six A2HD patients. (D) Correlation between the level of Tax11-19 peptide–HLA-A*201 expression and HTLV-I DNA load, RNA load, and HTLV-I Tax–specific CD8+ T cell frequency. The levels of Tax11-19 peptide–HLA-A*201 expression on CD4+ T cells were significantly correlated with HTLV-I proviral DNA load (P = 0.0490, r2 = 0.661), HTLV-I mRNA load (P = 0.0259, r2 = 0.749), and HTLV-I Tax11-19 tetramer–specific CD8+ T cell frequencies (P = 0.0470, r2 = 0.668) in six HAM/TSP patients.
Mentions: The inability to detect Tax11-19 peptide–HLA-A*201 complexes directly from ex vivo T cells of HAM/TSP patients (Fig. 2 A) could be reflective of a low concentration of Tax11-19 peptide bound to endogenous HLA-A*201 molecules. As we had shown that the staining intensity of the TaxA2-Ab was dependent on the concentration of the peptide used for pulsing T cells in A2HD (Fig. 1 F), it was of interest to determine whether this TaxA2-Ab staining of peptide-pulsed ex vivo T cells was different between A2HD and HLA-A*201 HAM/TSP patients. CD4+ T cells were separated from ex vivo peripheral blood T cells of six A2HD and six HLA-A*201 HAM/TSP patients. These ex vivo T cells were pulsed at a concentration of 10 μM Tax11-19 peptide, previously determined to be in a range where the TaxA2-Ab was unable to detect Tax11-19 peptide–HLA-A*201 complexes (Fig. 1 F). In contrast to HTLV-I Tax11-19 peptide–pulsed A2HD T cells (Figs. 5 A and 1 F), staining intensity of the TaxA2-Ab on comparably peptide-pulsed HAM/TSP T cells was significantly higher (Fig. 5, A and B). To determine if this difference between A2HD and HAM/TSP patients was specific for the amount of Tax11-19 peptide–HLA-A*201 complexes as defined by the TaxA2-Ab, CD4+ T cells from the same individuals were pulsed with the same concentration of control melanoma gp100 G9-154 peptide, and the level of G9-154 peptide–HLA-A*201 complexes was analyzed using the MelanomaA2-Ab. As shown in Fig. 5 A, staining intensity with the MelanomaA2-Ab in HAM/TSP patients was similar to that in A2HD. These observations were confirmed using a wider range of peptide concentrations (Fig. 6, A and B). As the staining sensitivity of the TaxA2-Ab might be affected by the amount of expressed HLA-A*201 complexes, we compared the amount of HLA-A*201 expression on CD4+ T cells between A2HD and HAM/TSP using anti–HLA-A*201–specific Ab (BB7.2). There was no statistically significant difference in the amount of HLA-A*201 expression between A2HD and HAM/TSP (Fig. 5 C), suggesting that differential levels of HLA-A*201 expression do not contribute to the TaxA2-Ab staining intensity between A2HD and HAM/TSP patients. In addition, we were able to identify two HLA-A*0201 asymptomatic carriers from which sufficient cells were available to analyze with the TaxA2-Ab. TaxA2-Ab staining intensities on CD4+ T cells of these two HLA-A*0201 asymptomatic carriers were similar with those on A2HD (unpublished data).

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