Papers by Kelly Cristina Gomes Manfrere
Scientific Reports, May 11, 2016
Individuals who remain HIV-seronegative despite repeated unprotected exposure to the virus are de... more Individuals who remain HIV-seronegative despite repeated unprotected exposure to the virus are defined as exposed seronegative (ESN) individuals. Innate and adaptive immunity, as well as genetic factors, provide ESNs with important advantages that allow for low infection susceptibility. The majority of HIV-1-infected individuals undergo antiretroviral therapy, which can decrease the level of HIV-1 exposure in ESNs. We analyzed type I interferon (IFN)-related antiviral and regulatory factors in peripheral blood mononuclear cells (PBMCs) and oral epithelial cells from serodiscordant couples. Our findings revealed that ESNs did not induce the expression of antiviral factors (APOBEC-3G, TRIM5-α, SAMDH1, STING, TBk1) or regulatory factors (Trex, Foxo3, Socs3, IL-10) in PBMCs, unlike their HIV-1infected partners. In contrast, ESNs upregulated APOBEC-3G and type I/III IFNs (IFNs-α,-β/-λ) in oral mucosal epithelial cells similar to their HIV-infected partners. The serodiscordant groups exhibited an increased expression of type I IFN-induced regulators, such as Trex and Foxo3, in oral epithelial cells. TLR7, TLR8 and TLR9 were expressed in oral epithelial cells of both ESNs and HIV-1-infected subjects. These findings revealed evidence of antiviral factors, type I/III interferon and regulatory factor expression only in the oral mucosal compartment of ESNs, while HIV-1-infected partners systemically and oral mucosal expressed the antiviral profile. High-risk populations (e.g., sex workers, serodiscordant couples, intravenous drug users, and children born from HIV-1-infected mothers) have been studied as exposed seronegative (ESN) individuals 1-4. The mechanisms involved in the protection against HIV infection are multifactorial, and the combined contributions of innate and adaptive immunity, as well as genetic factors, provide important advantages that allow for a low susceptibility to infection 5-7. Mucosal and systemic HIV-1-specific cellular and humoral responses may play defining roles in the protection against HIV-1 infection 8-12. By analyzing the innate immunity of ESNs, we verified that natural killer (NK) cells presented a unique activation profile, with increased levels of NKG2D, CD107a, and interferon (IFN)-γ expression and memory CD57 + CD56 dim NK cells 13. We also found that ESNs contained populations of Tc22/Th22 cells and polyfunctional staphylococcal enterotoxin B-induced CD4 + and CD8 + T cells with a low activation profile 14. Moreover, a dysfunctional innate immune response to Toll-like receptor (TLR) stimulation has been detected in HIV-1-infected mothers treated with antiretroviral therapy (ART) and their newborns 15. We also observed a significant increase in mRNA expression of antiviral factors in mononuclear cells from HIV-infected mothers and cord blood compared to uninfected mother-newborn pairs, including the apolipoprotein B mRNA-editing enzyme 3G (A3G), A3F, tripartite motif family-5α (TRIM-5α), TRIM-22, myxovirus resistance protein A (MxA), stimulator of IFN genes (STING) and IFN-β 16. HIV replication is limited by cellular restriction factors, such as the citidine deaminase A3G, which acts by inducing G to A mutations 17. ESNs expressed higher levels of A3G than healthy controls, suggesting that exposure to
Placenta, Apr 1, 2015
The involvement of angiotensin II (Ang II) in maximal decidualization has been reported. We demon... more The involvement of angiotensin II (Ang II) in maximal decidualization has been reported. We demonstrated that Ang II activates the Ca 2+ / calcineurin (CN)/ NFAT pathway in rat endometrial stromal cells (ESCs). Ang II modulates gene expression, proliferation and metalloprotease activity in ESCs that could be involved in decidualization. Objective: To analyze in vitro a) whether Ang II induces decidual reaction and uterine receptivity in ESCs, b) the involvement of calcineurin, and c) whether Ang II secreted by trophoblasts could mediate those effects. Methods: ESCs from rat uterus were cultured with Ang II or estrogen plus medroxyprogesterone acetate (E2+MPA), for 24 h or 6 days. ESCs were pretreated with Cyclosporin A to assess the involvement of CN. mRNA expression of desmin (decidual prolactin related protein (dprp)), and insulin growth factor binding protein 1 (Igfbp1), markers of decidualization, was measured by RT-PCR. mRNA and protein expression of HB-EGF and IGF-1, markers of uterine receptivity, were measured by RT-PCR and western blot. The rat choriocarcinoma-derived cell line Rcho-1, which can be differentiated in vitro into invasive trophoblasts, was cultured in the presence or absence of E2 (10-8 M) or MPA (1 mM) for 48 h. ESCs were then cultured with the trophoblast conditioned medium (TrCM) for 48 h in the presence or absence of the Ang II receptor antagonist Losartan. Results: Ang II induced (p<0.05) the expression of desmin, dprp, igfbp1, HB-EGF and IGF-1 in ESCs. Cyclosporin A inhibited (p<0.05) the effect of Ang II. MPA-TrCM, compared to control TrCM, induced dprp (1.7 fold) and HB-EGF (2.1 fold) expression in ESCs. Losartan inhibited TrCM induction (p<0.05). Conclusions: These results suggest that 1) the induction of markers of decidualization and uterine receptivity by Ang II appears to require calcineurin activity, 2) the activation of the renin-angiotensin-system in trophoblasts may induce the decidual reaction in ESCs.
Anais Brasileiros de Dermatologia
Primary cutaneous T-cell lymphomas (CTCL) are a heterogeneous group of non-Hodgkin lymphomas that... more Primary cutaneous T-cell lymphomas (CTCL) are a heterogeneous group of non-Hodgkin lymphomas that initially manifest in the skin. Sézary syndrome (SS) is a rare variant with an aggressive prognosis, characterized by the triad of erythroderma, lymphadenopathy and leukemic involvement. The malignant cell, called Sézary cells, is the TCD4+ lymphocyte, in the peripheral blood, with cerebriform nuclei. The contribution of inflammation, through inflammasomes, especially at the nodal site, is not yet known in patients with SS. The inflammasome is a multiprotein complex present in the cytosol, it is considered a receptor of molecular patterns which after stimulation culminates in the production of pro-inflammatory cytokines (IL-1β and IL-18) and pyroptosis. Our proposal was to evaluate the relationship between chronic inflammation by inflammasome components NLRP1, NLRP3, NLRP4, AIM-2 and cytokines (IL-1β/IL-18) in lymph node, skin and peripheral blood samples from SS. We selected 26 patients with SS (13 men, 13 women) aged 22-84 years from the Cutaneous Lymphoma Outpatient Clinic of the Hospital das Clínicas da Faculdade de Medicina da USP, and a control group (CTRL) with 28 healthy individuals (14 men, 14 women) aged between 30-85 years. For skin and lymph node analyses, patients with idiopathic erythroderma (IE) were used as a control group. In lymph nodes, protein expression analysis by immunohistochemistry indicates a reduction in the expression of IL-1β and an increase in the expression of IL-18 in the N2/N3 stages compared to the IE group, the presence of IL-18 was mainly in macrophages (CD68+). Furthermore, the increase in IL-18 transcript level was confirmed in circulating mononuclear cells and at the protein level in serum. The reduction in the expression of the IL-1β transcript and of IL-1B-related gene pathways was also confirmed by transcriptome analysis in the most severe stages compared to cases of IE lymph nodes. Interestingly, epidermal IL-1β was increased while IL-18 was decreased in the skin of patients with SS, in the dermal layer the expression of IL-18 was increased in relation to the IE group. The results show that in Sezary syndrome, the upregulation of IL-18 by macrophages can contribute to angiogenesis and favor the growth of tumor cells, and the inhibition of IL-1β could be a tumor escape preventing pyroptosis. These data may suggest new therapeutic targets for LCCT.
International Journal of Molecular Sciences
Sézary syndrome (SS) is a rare and aggressive type of cutaneous T-cell lymphoma, with an abnormal... more Sézary syndrome (SS) is a rare and aggressive type of cutaneous T-cell lymphoma, with an abnormal inflammatory response in affected skin. The cytokines IL-1B and IL-18, as key signaling molecules in the immune system, are produced in an inactive form and cleave to the active form by inflammasomes. In this study, we assessed the skin, serum, peripheral mononuclear blood cell (PBMC) and lymph-node samples of SS patients and control groups (healthy donors (HDs) and idiopathic erythroderma (IE) nodes) to investigate the inflammatory markers IL-1B and IL-18 at the protein and transcript expression levels, as potential markers of inflammasome activation. Our findings showed increased IL-1B and decreased IL-18 protein expression in the epidermis of SS patients; however, in the dermis layer, we detected increased IL-18 protein expression. In the lymph nodes of SS patients at advanced stages of the disease (N2/N3), we also detected an enhancement of IL-18 and a downregulation of IL-1B at the...
International Journal of Dermatology, 2020
Kevin Phan , MD Varitsara Mangkorntongsakul *, Bmed Vignesh Ramachandran , BS Asad Loya , BS Saxo... more Kevin Phan , MD Varitsara Mangkorntongsakul *, Bmed Vignesh Ramachandran , BS Asad Loya , BS Saxon D. Smith , MBChB, PhD, FACD Department of Dermatology, Liverpool Hospital, Sydney, Australia University of New South Wales, Sydney, Australia Gosford Hospital, Central Coast Local Health District, Gosford, New South Wales, Australia Baylor College of Medicine, Houston, TX, USA The Dermatology & Skin Cancer Centre, St Leonards, Sydney, Australia *E-mail: [email protected]
Frontiers in Immunology, 2020
Innate immunity is one of the main protection mechanisms against viral infections, but how this s... more Innate immunity is one of the main protection mechanisms against viral infections, but how this system works at the maternal-fetal interface, especially during HIV infection, is still poorly known. In this study, we investigated the relationship between pregnancy and innate mechanisms associated with HIV immunity by evaluating the expression of DAMPs, inflammasome components and type I/III IFNs in placenta and serum samples from HIV-infected mothers and exposed newborns. Our results showed that most of these factors, including HMGB1, IL-1, and IFN, were increased in placental villi from HIV-infected mothers. Curiously, however, these factors were simultaneously repressed in serum from HIV-infected mothers and their exposed newborns, suggesting that pregnancy could restrict HIV immune activation systemically but preserve the immune response at the placental level. An effective local antiviral status associated with a suppressed inflammatory environment can balance the maternal immune response, promoting homeostasis for fetal development and protection against HIV infection in neonates.
European Journal of Cancer, 2019
Oncotarget, Jan 9, 2018
Sézary syndrome (SS) is a leukemic variant of cutaneous T cell lymphoma (CTCL), and the neoplasti... more Sézary syndrome (SS) is a leukemic variant of cutaneous T cell lymphoma (CTCL), and the neoplastic CD4+ T cells of SS patients undergo intense clonal proliferation. Although Sézary cells have been studied extensively, studies on adaptive immunity regarding CD8+T cells are scarce. This study aimed to investigate activation marker expression in CD8+ T cells according to the differentiation stages and IL-7/IL7Rα axis responses of patients with SS. Moreover, this study aimed to verify the soluble forms of CD38, sCD127 and IL-7 in serum. Although the SS patients of our cohort had reduced numbers of CD8+ T cells, they exhibited higher percentages of CD8+CD38+ T cells, mainly effector/memory CD8+ T cells, than the control group. In contrast, down-regulated expression of the activation markers CD127/IL-7R and CD26 was found in the CD8+ T cells of SS patients. High serum levels of sCD38 and sCD127 and scarce serum levels of IL-7 were detected, emphasizing the immune activation status of SS p...
JAAD Case Reports, 2017
S ezary syndrome (SS) is a malignancy of CD4 1 central memory T-lymphocytes, characterized by the... more S ezary syndrome (SS) is a malignancy of CD4 1 central memory T-lymphocytes, characterized by the triad of erythroderma, lymphadenopathy, and involvement of peripheral blood by neoplastic cells. We describe a rare case of doublepositive CD4 and CD8 SS.
Oncotarget, 2017
Sézary syndrome (SS), an aggressive and leukemic form of cutaneous T-cell lymphoma, usually resul... more Sézary syndrome (SS), an aggressive and leukemic form of cutaneous T-cell lymphoma, usually results in shortened survival. Improving innate immunity in SS by targeting natural killer (NK) cells with Toll-like receptor (TLR) agonists could be an interesting modulatory strategy. We evaluated the NK cell populations in SS patients assessing activating and inhibitory receptors expression and profiled the differential expression of TLR signaling pathway genes in unstimulated NK cells and after TLR7/8 stimulation. We observed preserved CD56 bright NK cells and a low percentage of CD56 dim NK cells in the peripheral blood of SS patients compared to those in the healthy control group. Both NK cell populations showed down-modulation of NKG2C and NKG2D expression, which was associated with high serum levels of the soluble form of NKG2D ligands. In contrast, an expansion of "memory" CD57+ NKG2C+ NK cells and high cytomegalovirus antibody titers were detected in SS patients. Profiling of the TLR signaling genes in NK cells from SS patients showed an abundance of differentially expressed genes (DEGs) in NK cells in the unstimulated condition, with mostly upregulation of NFκB/JNK p38 pathway genes, but there was down-regulation of type I (IFN-α/β) and II (IFN-γ) interferon and IL-12A. After activation of NK cells with TLR7/8 agonist, the down-regulated genes correlated with the IFN response, and IL-12 became up-regulated, together with other antitumor factors. NK cell activation with a dual agonist for TLR7 and TLR8 is able to induce the expression of IFN-γ and type I IFN, which can improve immunity in SS patients.
Oncotarget, 2016
Sézary syndrome (SS) carries a poor prognosis, and infections represent the most frequent cause o... more Sézary syndrome (SS) carries a poor prognosis, and infections represent the most frequent cause of death in SS patients. Toll-like receptors (TLRs) are a family of innate immune receptors that induce protective immune responses against infections. We sought to evaluate the ability of TLR agonists to induce inflammatory cytokine, Th2 cytokine, and type I interferon (IFN-I) production by peripheral blood mononuclear cells (PBMC) of untreated SS patients. We detected impaired IL-6, IL-10 and IL-13 secretion by PBMC induced by the agonists for TLR5, TLR3, TLR7 and TLR9 in SS patients, while it was partially recovered by TLR2/TLR4 and TLR7/8 agonists TNF secretion was restored following stimulation with TLR2/TLR4 agonists. IFN-γ was scarcely produced upon TLR activation in SS cells, albeit TLR 7/8 (CL097) enhanced their secretion at lower levels than the control group. TLR9 agonist efficiently induced IFN-I in SS patients, although this positive regulation was not observed for other cytokines, in direct contrast to the broad activity of CL097. Among the TLR agonists, TLR4 was able to induce pro-inflammatory, IL-10 and Th2 secretion, while TLR7-8 agonist induced the inflammatory cytokines, IFN-I and IFN-γ. These findings reveal a dysfunctional cytokine response upon both extracellular and intracellular TLR activation in SS patients, which was partially restored by TLRs agonists. ACKNOWLEDGMENTS We are grateful to all the individuals who participated in the study.
Acta dermato-venereologica, Jan 3, 2015
Lichen planus (LP) is a chronic inflammatory mucocutaneous disease. The inflammatory status of LP... more Lichen planus (LP) is a chronic inflammatory mucocutaneous disease. The inflammatory status of LP may be related to S100A8 (myeloid-related protein 8; MRP8) activation of cytotoxic cells. The aims of this study were to evaluate S100A8 expression in skin lesions and the in vitro effects of S100A8 on CD8+ T cells and natural killer (NK) cells in LP. Increased levels of S100A8/S100A9 expression were measured in the skin lesions and sera of subjects with LP. S100A8 expression induced a greater cytotoxic response by peripheral blood CD8+CD107a+ T cells as well as by NK CD56bright cells in patients with LP. Increased expression of interleukin (IL)-1β, tumour necrosis factor (TNF) and IL-6 in the CD8+ T cells of patients with LP was induced by S100A8, in contrast to IL-10 and interferon (IFN) type I genes in the control group. These data suggest that, in individuals with LP, S100A8 may exert distinct immunomodulatory and cytotoxicity functions.
Ao Meu Deus, pois sem Ele eu não estaria aqui. À minha orientadora, Professora Dra Maria Notomi S... more Ao Meu Deus, pois sem Ele eu não estaria aqui. À minha orientadora, Professora Dra Maria Notomi Sato, pela orientação, carinho, paciência e por gerar em mim o amor pela ciência. Ao professor Dr Jose Antonio Sanches pela co-orientação, confiança, paciência e incentivos de que podemos melhorar a vida desses pacientes. À minha querida amiga e parceira de trabalho Marina Passos Torrealba, pelo apoio, carinho, comprometimento com nosso trabalho. Ao Dr Denis Ricardo Myashiro pela ajuda, de grande valia, na elaboração da dissertação e desenvolvimento do projeto. Àamiga de coração Nátalli Zanette pelo apoio e explicações ilustradas em rascunhos que me fez entender além dos livros. Às amizades nascidas no laboratório que ajudaram a manter a calma, a seguir a diante mesmo nas horas de desânimo, que pacientemente dividiam seus conhecimentos práticos, teóricos e pessoais e por todos os momentos de convívio e companheirismo:
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Papers by Kelly Cristina Gomes Manfrere