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    Tumor hypoxia shapes natural killer cell anticancer activities
    (Springer Science and Business Media LLC, 2025-05-30)
    RETAMAL LUCERO, MAURICIO ANTONIO  
    ;
    Flavio Salazar-Onfray
    ;
    Fermín E. González
    ;
    Andrés Tittarelli
      1
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    TGFβ links EBV to multisystem inflammatory syndrome in children
    (Springer Science and Business Media LLC, 2025-03-12)
    Carl Christoph Goetzke
    ;
    Mona Massoud
    ;
    Stefan Frischbutter
    ;
    Gabriela Maria Guerra
    ;
    Marta Ferreira-Gomes
    <jats:title>Abstract</jats:title> <jats:p>In a subset of children and adolescents, SARS-CoV-2 infection induces a severe acute hyperinflammatory shock<jats:sup>1</jats:sup> termed multisystem inflammatory syndrome in children (MIS-C) at four to eight weeks after infection. MIS-C is characterized by a specific T cell expansion<jats:sup>2</jats:sup> and systemic hyperinflammation<jats:sup>3</jats:sup>. The pathogenesis of MIS-C remains largely unknown. Here we show that acute MIS-C is characterized by impaired reactivation of virus-reactive memory T cells, which depends on increased serum levels of the cytokine TGFβ resembling those that occur during severe COVID-19 (refs. <jats:sup>4,5</jats:sup>). This functional impairment in T cell reactivity is accompanied by the presence of TGFβ-response signatures in T cells, B cells and monocytes along with reduced antigen-presentation capabilities of monocytes, and can be reversed by blocking TGFβ. Furthermore, T cell receptor repertoires of patients with MIS-C exhibit expansion of T cells expressing TCRVβ21.3, resembling Epstein–Barr virus (EBV)-reactive T cell clones capable of eliminating EBV-infected B cells. Additionally, serum TGFβ in patients with MIS-C can trigger EBV reactivation, which is reversible with TGFβ blockade. Clinically, the TGFβ-induced defect in T cell reactivity correlates with a higher EBV seroprevalence in patients with MIS-C compared with age-matched controls, along with the occurrence of EBV reactivation. Our findings establish a connection between SARS-CoV-2 infection and COVID-19 sequelae in children, in which impaired T cell cytotoxicity triggered by TGFβ overproduction leads to EBV reactivation and subsequent hyperinflammation.</jats:p>
    Scopus© Citations 4  2
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    Clinical, immunologic, and genetic characteristics of 148 patients with natural killer cell deficiency
    (Elsevier BV, 2025-05)
    Manar Abdalgani
    ;
    Evelyn R. Hernandez
    ;
    Luis A. Pedroza
    ;
    Ivan K. Chinn
    ;
    Lisa R. Forbes Satter
    Scopus© Citations 1  2
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      2Scopus© Citations 13
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    Partial loss-of-function mutations in GINS4 lead to NK cell deficiency with neutropenia
    (2022)
    Matilde I. Conte
    ;
    M. Cecilia Poli
    ;
    Angelo Taglialatela
    ;
    Giuseppe Leuzzi
    ;
    Ivan K. Chinn
      9Scopus© Citations 13
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    Improving Cell Recovery: Freezing and Thawing Optimization of Induced Pluripotent Stem Cells
    (2022)
    Markus Uhrig
    ;
    EZQUER, EDUARDO FERNANDO  
    ;
    EZQUER, EDUARDO MARCELO  
    <jats:p>Achieving good cell recovery after cryopreservation is an essential process when working with induced pluripotent stem cells (iPSC). Optimized freezing and thawing methods are required for good cell attachment and survival. In this review, we concentrate on these two aspects, freezing and thawing, but also discuss further factors influencing cell recovery such as cell storage and transport. Whenever a problem occurs during the thawing process of iPSC, it is initially not clear what it is caused by, because there are many factors involved that can contribute to insufficient cell recovery. Thawing problems can usually be solved more quickly when a certain order of steps to be taken is followed. Under optimized conditions, iPSC should be ready for further experiments approximately 4–7 days after thawing and seeding. However, if the freezing and thawing protocols are not optimized, this time can increase up to 2–3 weeks, complicating any further experiments. Here, we suggest optimization steps and troubleshooting options for the freezing, thawing, and seeding of iPSC on feeder-free, Matrigel™-coated, cell culture plates whenever iPSC cannot be recovered in sufficient quality. This review applies to two-dimensional (2D) monolayer cell culture and to iPSC, passaged, frozen, and thawed as cell aggregates (clumps). Furthermore, we discuss usually less well-described factors such as the cell growth phase before freezing and the prevention of osmotic shock during thawing.</jats:p>
      6Scopus© Citations 34
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    Surface Immunogenic Protein of Streptococcus Group B is an Agonist of Toll-Like Receptors 2 and 4 and a Potential Immune Adjuvant
    (2020)
    Diego A. Diaz-Dinamarca
    ;
    Ricardo A. Manzo
    ;
    Daniel A. Soto
    ;
    María José Avendaño-Valenzuela
    ;
    Diego N. Bastias
    <jats:p>Vaccine-induced protection against pathogens, especially subunit-based vaccines, are related to antigen properties but mainly in their ability to stimulate the immune system by the use of an adjuvant. Modern vaccines are formulated with a high level of antigen purity, where an efficient adjuvant is necessary. In this context, the use of protein Toll-Like Receptor (TLR) agonists as vaccine adjuvants has been highlighted because of their optimal immunogenicity and minimal toxicity. The Surface Immunogenic Protein (SIP) from Group B Streptococcus (GBS) has gained importance as a new potential protein-based vaccine. Recently, we reported that recombinant SIP (rSIP) expressed by E. coli and purified by High Performance Liquid Chromatography (HPLC) alone induces a protective humoral immune response. In this study, we present the immunomodulatory properties of rSIP as a protein-based adjuvant, as an agonist of TLR. To this end, we showed that C57BL/6 bone marrow-derived dendritic cells pulsed by rSIP resulted in enhanced CD40, CD80, CD86, and Major Histocompatibility Complex (MHC) class II as well as increased secretion proinflammatory cytokines Interleukin (IL)-6, Interferon (IFN)-γ, Tumor Necrosis Factor (TNF)-α, and IL-10. Next, we investigated the in vivo effect of rSIP in the absence or presence of ovalbumin (OVA) on antigen-specific antibody secretion in C57BL/6 mice. Immunization with rSIP plus OVA showed that anti-OVA IgG2a and IgG1a increased significantly compared with OVA alone in C57BL/6 mice. Also, the immunization of rSIP plus OVA generates increased serum cytokines levels characterized by IL-12p70, IL-10, IL-4, and IFN-γ. Interestingly, we observed that rSIP stimulate Toll Like Receptor (TLR)2 and TLR4, individually expressed by Human embryonic kidney (HEK) 293-derived TLR reporter cells. These findings suggest that rSIP is a new potential protein TLR agonist adjuvant and may be employed in the development of new vaccines.</jats:p>
      7Scopus© Citations 8
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    Potential use of n-3 PUFAs to prevent oxidative stress-derived ototoxicity caused by platinum-based chemotherapy
    (2020)
    Ignacio A. Cortés Fuentes
    ;
    Mauricio Burotto
    ;
    Mauricio A. Retamal  
    ;
    Michael Frelinghuysen
    ;
    Christian Caglevic
    Scopus© Citations 4  1
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    Safety of the intravenous administration of neurotensin-polyplex nanoparticles in BALB/c mice
    (2014)
    Maria E. Hernandez
    ;
    Jesus D. Rembao
    ;
    Daniel Hernandez-Baltazar
    ;
    Rosa A. Castillo-Rodriguez
    ;
    Victor M. Tellez-Lopez
      10Scopus© Citations 28
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    Epanorin, a lichen secondary metabolite, inhibits proliferation of MCF-7 breast cancer cells.
    (2019)
    Juan Palacios-Moreno
    ;
    Cecilia Rubio
    ;
    Wanda Quilhot
    ;
    M. Fernanda Cavieres
    ;
    Eduardo de la Peña
    <jats:title>Abstract</jats:title> <jats:sec> <jats:title>Background</jats:title> <jats:p>Epanorin (EP) is a secondary metabolite of the <jats:italic>Acarospora</jats:italic> lichenic species. EP has been found in lichenic extracts with antimicrobial activity, and UV-absorption properties have been described for closely related molecules; however, its antiproliferative activity in cancer cells has not yet been explored. It has been hypothesized that EP inhibits cancer cell growth. MCF-7 breast cancer cells, normal fibroblasts, and the non-transformed HEK-293 cell line were exposed to increasing concentrations of EP, and proliferation was assessed by the sulforhodamine-B assay.</jats:p> </jats:sec> <jats:sec> <jats:title>Results</jats:title> <jats:p>MCF-7 cells exposed to EP were examined for cell cycle progression using flow cytometry, and DNA fragmentation was examined using the TUNEL assay. In addition, EP’s mutagenic activity was assessed using the <jats:italic>Salmonella typhimurium</jats:italic> reverse mutation assay. The data showed that EP inhibits proliferation of MCF-7 cells, and it induces cell cycle arrest in G0/G1 through a DNA fragmentation-independent mechanism. Furthermore, EP’s lack of overt cytotoxicity in the normal cell line HEK-293 and human fibroblasts in cell culture is supported by the absence of mutagenic activity of EP.</jats:p> </jats:sec> <jats:sec> <jats:title>Conclusion</jats:title> <jats:p>EP emerges as a suitable molecule for further studies as a potential antineoplastic agent.</jats:p> </jats:sec>
    Scopus© Citations 7  20