Data_Sheet_1_Dermatological concerns for women and girls with turner syndrome.PDF

IntroductionTurner syndrome (TS) is associated with distinct manifestations in women and girls including short stature, cardiac abnormalities, premature ovarian failure as well as dermatological features, including lymphedema, keloids, onychodystrophy, and acne. Although many dermatological concerns present during the first few decades of life, the overwhelming majority of respondents are not provided with dermatology referrals at diagnosis.MethodsThis cross-sectional study utilized an author designed survey to assess self-reported dermatological manifestations, dermatology referral experience, common therapies for select dermatological conditions, as well as a validated 10-question Dermatology Life Quality Index (DLQI) to assess quality-of-life impact in women and girls with Turner syndrome.ResultsIn our cohort, 64% (n = 149) had been referred to a dermatologist at some point in their life time. The majority of individuals self-identified their dermatological concern (79.6%) and were referred after a dermatological concern had already occurred (90.2%). The most common dermatological findings reported were xerosis cutis (78.7%), lymphedema (73%), and more than 20 acquired melanocytic nevi (70%). The overall mean DLQI score was 3.52, indicative of a small effect on the patient’s life. Onychodystrophy, history of skin biopsy, and lymphedema were statistically significant to have a higher impact on quality of life.DiscussionOur data reveal that skin conditions are highly prevalent in the TS population during the early decades of life and affirm utilizing these conditions in the TS diagnostic process, as well as emphasize the need for specialized dermatology referrals to address the detrimental impacts related to skin concerns on quality of life.</p

The Role of IL-12 Signaling in Enhanced Anti-HIV Immunity

<div><p>(A and B) In vivo injection with IL-12 preferentially enhanced gp120-specific CTL and Th responses induced by SOCS1-silenced DCs. C57BL/6 mice were immunized with 1 × 10⁶ of HIV gp120-pulsed (50 μg/ml), transduced DCs derived from BM of WT mice or IL-12 receptor KO mice with ex vivo TNFα maturation (50 ng/ml). On days 1, 3, and 5 after DC immunization, murine IL-12 (1 μg/mouse, Peprotech) was administered intraperitoneally. CD8⁺ T cells (A) or CD4⁺ T cells (B) isolated 2 wk later from the pooled splenocytes of immunized mice (2–3 each group) were subjected to IFN-γ ELISPOT assays. An irrelevant protein, OVA, was used as a negative control. Representative data from two independent experiments are presented. *<i>P</i> < 0.01, LV-SOCS1-siRNA-DC versus LV-SOCS1-siRNA-DC + IL-12, or IL12R KO LV-SOCS1-siRNA-DC + IL-12 versus LV-SOCS1-siRNA-DC + IL-12.</p> <p>(C and D) gp120-specific CTL and Th responses induced by SOCS1-silenced DCs or Ad-IL-12-DCs. BM-derived DCs from WT mice were transfected with LV-SOCS1-siRNA (MOI of 5) or Ad-IL-12 with various MOIs of 10–1,000 or cotransfected with LV-SOCS1-siRNA (MOI of 5) and Ad-IL-12 (MOI of 10) for 4 h. DCs derived from BM of IL-12 receptor KO mice were cotransfected with LV-SOCS1-siRNA (MOI of 5) and Ad-IL-12 (MOI of 10) for 4 h. Groups of C57BL/6 mice were immunized with 1 × 10⁶ of gp120-pulsed (50 μg/ml), transfected DCs with ex vivo TNFα maturation. CD8⁺ T-cells (C) or CD4⁺ T cells (D) isolated 2 wk later from the pooled splenocytes of immunized mice (2–3 each group) were subjected to IFN-γ ELISPOT assays. An irrelevant protein, OVA, was used as a negative control. Representative data from two independent experiments are presented. <i>P</i> < 0.01, Ad-IL-12/SOCS1-siRNA-DC versus IL-12-DCs, or Ad-IL-12/SOCS1-siRNA-DC versus IL12R KO Ad-IL-12/SOCS1-siRNA-DC.</p></div

Enhanced gp120-Specific Antibody and T Cell Responses Induced by SOCS1-Silenced DCs

<p>Groups of C57BL/6 mice were immunized with gp120 (SF162) protein-pulsed, transduced BM-derived DCs (1 × 10⁶ cells/mouse) twice at a weekly interval, followed by PolyI:C or R837 stimulation (30 μg/mouse) in vivo three times on days 1, 3, and 5 after each DC immunization, and sera and splenocytes were collected from each group of mice 14 d later. HIV gp120-specific IgG subclass titers (A) from the pooled sera of each group (4–6 mice/group) were quantified by capture ELISA. CD8⁺ T cells (B) and CD4⁺ T cells (C) isolated from pooled splenocytes were used for IFN-γ ELISPOT assays stimulated with gp120 proteins. Intracellular IFN-γ staining of CD8⁺ T cells from the pooled splenocytes were also performed (D). Representative data from one of three experiments are presented. NS, no stimulation. *<i>P</i> < 0.01, LV-SOCS1-siRNA-DCs versus LV-GFP-siRNA-DCs.</p

Resistance of SOCS1-Silenced DCs to HIV gp120-Mediated Suppression

<div><p>(A) Effects of gp120 on cytokine production by DCs. BM-DCs were transfected with SOCS1-siRNA or a SOCS1-siRNA mutant oligonucleotide as described previously [<a href="http://www.plosmedicine.org/article/info:doi/10.1371/journal.pmed.0030011#pmed-0030011-b024" target="_blank">24</a>], and then cultured with or without SF162 gp120 (20 μg/ml) or LPS (100 ng/ml), and cytokine levels were determined at the different times of cultures, as indicated.</p> <p>(B–E) Effects of gp120 on DC antigen presentation in vivo. Transfected BM-DCs were pulsed with OVA, incubated with or without gp120 for 2 d, and then stimulated with LPS (100 ng/ml) ex vivo overnight. Mice were then immunized with the transduced DCs twice, following three in vivo LPS stimulations. OVA-specific antibody IgG (B) and IgG1 (C) titers and frequencies of IFN-γ-producing OVA-specific CD8⁺ T cells (D) and CD4⁺ T cells (E) were examined 2 wk after the second DC immunization. Data are representative of two repeats.</p></div

Enhanced Production of Both Th1- and Th2-Polarizing Cytokines by SOCS1-Silenced DCs and Activated CD4⁺ Th

<div><p>(A) Enhanced production of both Th1- and Th2-polarizing cytokines by SOCS1-silenced DCs. BM-DCs transfected with SOCS1 siRNA or control [<a href="http://www.plosmedicine.org/article/info:doi/10.1371/journal.pmed.0030011#pmed-0030011-b024" target="_blank">24</a>] were stimulated with LPS (100 ng/ml). Concentrations of various cytokines in the culture media were analyzed by ELISA 24 h after stimulation. NS, no stimulation. *<i>P</i> < 0.01, LV-SOCS1-siRNA-DCs versus LV-GFP-siRNA-DCs.</p> <p>(B) IL-12 production by transduced DCs. BM cells were cultured with mGM-CSF (20 μg/ml only or mGM-CSF and mIL-4 (20 μg/ml) [<a href="http://www.plosmedicine.org/article/info:doi/10.1371/journal.pmed.0030011#pmed-0030011-b024" target="_blank">24</a>] for 6 d and then transduced with LV-SOCS1-siRNA or LV-GFP-siRNA. The transduced DCs (5 × 10⁵/ml) were then stimulated with LPS (100 ng/ml) and plate-coated anti-CD40 mAb (5 μg/ml, BD Bioscience) in the presence or absence of IL-4. Concentrations of IL-12 (p70) in the culture media were analyzed by ELISA 24 h after stimulation and are presented from one of three independent experiments. *<i>P</i> < 0.01, LV-SOCS1-siRNA (GM-CSF + IL-4) versus LV-GFP-siRNA (GM-CSF + IL-4).</p> <p>(C–E) CD4⁺ Th responses induced by LV-SOCS1-siRNA-DCs. CD4⁺ T cells were isolated from pooled splenocytes of different groups of mice and subjected to the following assays. Numbers of IFN-γ-producing CD4⁺ T cell precursors were determined with the ELISPOT assay (C). ³H-thymidine incorporation rates of the isolated CD4⁺ T cells were determined on the fourth day of restimulation with gp120-pulsed DCs (D). Cytokine levels in the culture medium of isolated CD4⁺ cells stimulated with gp120-pulsed DCs for 48 h were determined by ELISA (E). The mean results (+ standard error) from one of three experiments are presented. *<i>P</i> < 0.01, LV-SOCS1-siRNA-DC versus LV-GFP-siRNA-DC mice.</p></div

Long-Term gp120-Specific Antibody and CTL Responses Induced by SOCS1-Silenced DCs

<p>IgG subclass titers (A) from pooled sera of different groups of mice and frequencies of IFN-γ-positive T cells of CD8⁺ T cells (B) and CD4⁺ T cells (D) isolated from pooled splenocytes (two mice per group) were determined at 6 mo after DC immunization and on day 7 after booster immunization with recombinant gp120 emulsified in IFA (20 μg protein/mouse). Intracellular IFN-γ and surface CD44 costaining of gated CD8⁺ T cells from splenocytes at 6 mo after immunization are shown (C). The data are representative of two experiments.</p

Enhancement of HIV DNA Vaccine by SOCS1-siRNA DNA

<p>Groups of C57BL/6 mice were immunized with gp140CF DNA [<a href="http://www.plosmedicine.org/article/info:doi/10.1371/journal.pmed.0030011#pmed-0030011-b074" target="_blank">74</a>] only or a mixture of gp140CF DNA and pSuper-SOCS1-siRNA expressor DNA [<a href="http://www.plosmedicine.org/article/info:doi/10.1371/journal.pmed.0030011#pmed-0030011-b024" target="_blank">24</a>] weekly for 3 wk, followed by PolyI:C or R837 stimulation (30 μg/mouse) in vivo three times on days 1, 3, and 5 after each DNA immunization. HIV gp120-specific IgG subclass titers (A), IFN-γ spot numbers of CD8⁺ T cells (B) and CD4⁺ T cells (C) from pooled splenocytes of different groups of mice (4–6 mice per group) 1 wk after the last immunization are shown from one of three independent experiments. Intracellular IFN-γ staining of CD8⁺ T cells from the pooled splenocytes was also performed (D). *<i>P</i> < 0.01, gp140CF and GFP siRNA versus gp140CF and SOCS1 siRNA coimmunization.</p

Enhanced HIV-Specific B Cell Responses

<div><p>(A) Enhanced production of BAFF and APRIL by SOCS1-silenced DCs. The transduced BM-DCs were stimulated with LPS (100 ng/ml) for 24 h. Relative expression levels of BAFF and APRIL mRNA were then determined by real-time quantitative PCR as described in Methods, and normalized to mock-transfected DCs after LPS stimulation using the Comparative Ct method [<a href="http://www.plosmedicine.org/article/info:doi/10.1371/journal.pmed.0030011#pmed-0030011-b027" target="_blank">27</a>]. Representative data from two independent experiments are presented. *<i>P</i> < 0.01, LV-SOCS1-siRNA-DC versus LV-GFP-siRNA-DCs.</p> <p>(B–D) Enhanced activation of gp120-specific B cells by SOCS1-silenced DCs. Frequencies of anti-gp120 antibody-producing cells in different groups of mice were determined and reported as the number of cells secreting gp120-specific IgG per 5 × 10⁵ B cells (B). The proliferation rates (C) and cytokine production (D) of B cells (5 × 10⁴/well) isolated from the spleens of different groups of mice after stimulation with anti-CD40 (5 μg/ml), IL-4 (20 ng/ml), or costimulation with anti-CD40 and IL-4 for 48 h were determined, and results from one of three independent experiments are presented. <i>P</i> < 0.01, LV-SOCS1-siRNA-DC mice versus LV-GFP-siRNA-DC mice under various in vitro stimulation conditions.</p></div

Comparison of gp120-Specific Antibody and T Cell Responses Induced by Protein Immunization and SOCS1-Silenced DCs

<p>Groups of C57BL/6 mice were immunized with gp120 protein (20 μg/ml)-pulsed, transduced BM-derived DCs (1 × 10⁶ cells/mouse) or the same amount of gp120 protein formulated in IFA (20 μg/mouse) twice at a weekly interval. All of the mice were injected with PolyI:C or R837 (30 μg/mouse) in vivo three times on days 1, 3, and 5 after each immunization, and sera and splenocytes were collected from each group of mice 14 d later. HIV gp120-specific IgG subclass titers (A) from the pooled sera of each group (4–6 mice/group) were quantified by capture ELISA. CD8⁺ T cells (B) and CD4⁺ T cells (C) isolated from pooled splenocytes were used for IFN-γ ELISPOT assays stimulated with gp120 proteins. Representative data from one of three experiments are presented. *<i>P</i> < 0.01, gp120 protein + IFA versus gp120-pulsed LV-SOCS1-siRNA-DCs.</p