Immunological Profile of HTLV-1-Infected Patients Associated with Infectious or Autoimmune Dermatological Disorders

<div><p>In the present study, the frequency, the activation and the cytokine and chemokine profile of HTLV-1 carriers with or without dermatological lesions were thoroughly described and compared. The results indicated that HTLV-1-infected patients with dermatological lesions have distinct frequency and activation status when compared to asymptomatic carriers. Alterations in the CD4⁺HLA-DR⁺, CD8⁺ T cell, macrophage-like and NKT subsets as well as in the serum chemokines CCL5, CXCL8, CXCL9 and CXCL10 were observed in the HTLV-1-infected group with skin lesions. Additionally, HTLV-1 carriers with dermatological skin lesions showed more frequently high proviral load as compared to asymptomatic carriers. The elevated proviral load in HTLV-1 patients with infectious skin lesions correlated significantly with TNF-α/IL-10 ratio, while the same significant correlation was found for the IL-12/IL-10 ratio and the high proviral load in HTLV-1-infected patients with autoimmune skin lesions. All in all, these results suggest a distinct and unique immunological profile in the peripheral blood of HTLV-1-infected patients with skin disorders, and the different nature of skin lesion observed in these patients may be an outcome of a distinct unbalance of the systemic inflammatory response upon HTLV-1 infection.</p></div

Plasmatic levels of chemokines and cytokine ratios among HTLV-1-infected patients with skin lesions.

<p>Serum chemokines and cytokines quantitation were performed by CBA as described in <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0002328#s2" target="_blank">materials and methods</a>. The results are expressed as concentration in pg/mL (×10³) ± standard error of (A) CCL2, (B) CCL5, (C) CXCL8, (D) CXCL9, (E) CXCL10 in the serum of HTLV-infected patients and controls with infectious (L^(INF) = dark gray rectangles) and autoimmune (L^(AI) = black-gray rectangles) skin lesions as well as carriers and controls without skin lesions (L⁽⁻⁾ = light gray rectangles). Significant differences at <i>P</i><0.05 are highlighted by letters for difference of a group as compared to control L⁽⁻⁾ indicated by “a”, to control L^(INF) indicated by “b”, to control L^(AI) indicated by “c”, to HTLV-1+ L⁽⁻⁾ indicated by “d” and to HTLV-1+ L^(INF) indicated by “e”. The ratio between cytokines and IL-10 for all groups was calculated for (F) IL-1β, (G) IL-6, (H) IL-12, (I) TNF-α. Significant differences are highlighted by connecting lines.</p

Phenotypic profile of HTLV-1-infected patients and uninfected controls with skin lesions.

<p>Phenotypic studies were performed by a FACS double-labeling protocol as described in <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0002328#s2" target="_blank">materials and methods</a>. The results are expressed as mean percentage (%) ± standard error of (A) CD3⁺ T cells, (B) CD19⁺ B cells, (C) T/B ratio, (D) CD4⁺ T cells, (E) CD8⁺ T cells, (F) CD4⁺/CD8⁺ ratio, (G) CD4⁺HLA-DR⁺ (H) CD8⁺HLA-DR⁺ (I) CD4⁺HLA-DR⁺/CD8⁺HLA-DR⁺ in the peripheral blood from HTLV-1-infected patients and controls with infectious (L^(INF) = dark gray rectangles) and autoimmune (L^(AI) = black-gray rectangles) skin lesions as well as carriers and controls without skin lesions (L⁽⁻⁾ = light gray rectangles). Significant differences at <i>P</i><0.05 are expressed by connecting lines.</p

Proviral load of HTLV-1-infected patients without and with infectious or autoimmune skin lesions.

<p>(A) Proviral load was measured by Real Time PCR as described in <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0002328#s2" target="_blank">Materials and Methods</a> and results are expressed in number of HTLV-1 genome copies per 10⁴ peripheral blood mononuclear cells (PBMCs). The frequency of high (black rectangles #6 – 2 without skin lesions and 4 with skin lesions) medium (gray rectangles #11 – 1 without skin lesions and 10 with skin lesions) and low (white rectangles #19 – 6 without skin lesions and 13 with skin lesions) copy number was defined by the global arithmetic mean of all data (B) and individual groups (C).</p

Description of patients included in the cell subset analysis.

#<p>- Age average in years;</p>*<p>M/F – Male/Female ratio.</p

Immunoregulation in HTLV-1-infected patients with skin lesions of infectious or autoimmune nature.

<p>The schematic representation illustrates the cytokines, chemokines and cell populations altered in the peripheral blood of patients chronically infected by HTLV-1 with infectious or autoimmune skin lesions. These immunological features give insight to the understanding of how the immune systemic biomarkers may be involved in the generation of protective and pathogenic responses and their potential association with proviral load and skin lesion.</p

Phenotypic profile of innate immunity subsets.

<p>Monocyte subsets, NK-cells and NKT-cells and Regulatory T-cells in the peripheral blood of HTLV-1-infected patients and uninfected controls according to the absence (L⁽⁻⁾) or presence (L⁽⁺⁾) of dermatological Lesions.</p>*<p>Data are expressed as mean percentage (%) ± standard error of cell phenotype within a selected gated cell subset shown after the slash, except for CD3⁻CD16^−/+CD56^−/+ and CD4⁺CD25^HIGH that were analyzed within gated lymphocytes.</p><p>Significant differences at <i>P</i><0.05 are highlighted by letter “a”, “b” and “c” for difference between pairs of subgroups “All”, L⁽⁻⁾ and L⁽⁺⁾, respectively.</p

Description of patients included in the cytokine/chemokine analysis.

#<p>- Age average in years;</p>*<p>M/F – Male/Female ratio.</p

Frequency of high producers of chemokines and cytokines among HTLV-1-infected patients with skin lesions.

<p>The group with skin lesions was subdivided in the sub-groups of patients with infectious and autoimmune skin lesions. The frequency of high producers of chemokines (in gray) and cytokines (in black) is displayed in radar graphs with three stages (30% each) from 0 to 90% (as described in the scale located in the upper right corner of the graph) and it was calculated using the global arithmetic mean percentage of chemokines/cytokine levels as a threshold to define high and low producers.</p