Isabel Silva’s research while affiliated with Unidade Local de Saúde de Matosinhos and other places
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Introduction and objectives:
Patients with systemic autoimmune rheumatic disease (SARD) are a vulnerable population for severe COVID-19 and worse response to vaccination, prompting the need of a booster vaccine. Data regarding its response is limited and inconsistent. The aim of this study was to assess the effectiveness and immunogenicity of the third dose of the SARS-CoV-2 vaccine in immunosuppressed SARD patients.
Materials and methods:
We conducted a prospective study in immunosuppressed SARD Portuguese patients, who received a SARS-CoV-2 booster vaccine, from October 2021 to August 2022. We evaluated COVID-19 incidence in the following 6 months, as well as vaccine immunogenicity through anti-Spike IgG titers and T-cell reactivity to the Spike protein.
Results:
We included 131 patients with a mean age of 54.9±12.2 years. Almost 40% (n=52) developed COVID-19 within 6 months after the booster, but 51 (98.1%) were mild infections. Median post-booster antibody levels and antibody variation were 9540.7 (14,724) and 8937.9 (11,561.3)AU/mL, respectively, and 73.3% (n=96) of the patients showed post-booster T-cell reactivity. Antibody variation was significantly lower in the COVID group (p=0.015). Although post-booster antibody levels and T-cell reactivity were statistically significantly lower in the patients under biologic DMARD, there was not a significant increase in COVID-19 incidence.
Conclusions:
This study shows that a booster vaccine elicits strong immunogenicity and reduces COVID-19 severity, highlighting its importance in immunosuppressed SARD patients. Larger and more homogeneous cohorts are needed to guide periodic booster administration in this susceptible population.
Background: Breast cancer is a heterogeneous malignant disease with a varying prognosis and is classified into four molecular subtypes. It remains one of the most prevalent cancers globally, with the tumor microenvironment playing a critical role in disease progression and patient outcomes. Methods: This study evaluated tumor samples from 40 female patients with luminal A and B breast cancer, utilizing flow cytometry to phenotypically characterize the immune cells and tumor cells present within the tumor tissue. Results: The luminal B-like tumor samples exhibited increased infiltration of CD4⁺ cells, regulatory T cells (Tregs), and Th17 cells and decreased levels of NK cells, γδ T cells, Th1 cells, and follicular T cells, which is indicative of a more immunosuppressive tumor microenvironment. Conclusions: These findings suggest that luminal B-like tumors have a microenvironment that is less supportive of effective anti-tumor immune responses compared to luminal A tumors. This study enhances the understanding of the immunological differences between luminal subtypes of breast cancer and identifies potential new therapeutic targets and biomarkers that could drive advancements in precision medicine for breast cancer management.
A Doença Gestacional do Trofoblasto (DGT) é um conjunto heterogéneo de doenças raras relacionadas com o desenvolvimento anormal do trofoblasto na gravidez. Entre os fatores de risco associados destacam-se a idade materna avançada e antecedente de gestação molar, que variam de acordo com o subtipo de anomalia. A determinação do prognóstico, nas formas benignas, é feita pela monitorização dos valores de gonadotropina coriónica humana sanguínea (β-hCG), após tratamento. Nas formas malignas, é estabelecido pela correlação entre dois sistemas de classificação que permite categorizar os tumores em grupos de risco de resistência ao tratamento.
Introduction: The majority of Waldenstrom macroglobulinemia (WM) patients (pts) harbor the activating mutation MYD88L265P and about ⅓ nonsense or frameshift mutations in the CXCR4 gene. Both mutations have prognostic as well as therapeutic implications. B-cell malignancies present differences in the migratory and invasive patterns of malignant cells. Little is known about the mechanisms governing migratory heterogeneity, despite growing evidence concerning the role of homeostatic chemokines and their receptors, mainly CXCR4 and CXCR5, and adhesion molecules such as CD49d, in the dissemination pattern of lymphoid malignancies. Also, there are no data about its correlation with genetic characteristics of WM.
Aims: To evaluate the relationship between CXCR5, CXCR4 and CD49d protein expression levels in normal (nr) and abnormal (ab) lymphocytes and their role as potential surrogate markers for MYD88L265P and CXCR4 gene mutational status in WM.
Methods: We prospectively studied bone marrow aspirate samples of 45 pts newly diagnosed with WM from 3 Portuguese centers. MYD88L265P mutation testing performed in 39/45 (87%) cases by Real Time ASO-PCR and CXCR4 gene mutations by Sanger sequencing in 30/45 (67%) cases, preferably after sorting of CD19+ cells. Flow cytometric (FCM) expression of CXCR5, CXCR4 and CD49d evaluated in nr (normal k/l ratio) and ab B cells (monoclonal). FCM performed using a BD FACSCanto II cytometer. Data acquisition carried out using FACSDiva software and ≥10^3 events/sample were collected. Median fluorescence intensities (MFI) of the 3 molecules evaluated and a log2 transformation applied to all MFI values, to allow a more interpretable scale and to reduce skewness. MFI and ratios nr/ab (normalized variables) statistically compared across populations by Mann-Whitney U test.
Univariate logistic regression conducted to explore the association between the phenotypic markers and the genetic findings. Factors with p-values < 0.05 considered significant.
Results: We included 45 pts, median age 76 years, 57.8% male. Seventy-seven percent of pts were positive for MYD88L265P mutation and had no mutation in CXCR4 gene, 10% were positive for both and 13% were double negative. The median (min-max) of MFI was 11.83 (8.75-14.15) for CXCR5nr, 11.43 (8.71-14.24) for CXCR5ab, 10.50 (6.82-13.10) for CXCR4nr, 10.43 (7.48-13.63) for CXCR4ab, 9.95 (5.49-12.73) for CD49dnr and 9.83 (7.46-11.57) for CD49dab. There were no MFI differences between nr and ab lymphocytes in CXCR5 (p=0.233), CXCR4 (p=0.988) or CD49d (p=0.713). There was no correlation between CXCR5nr/CXCR4nr (r=-0.085; p=0.662), CXCR5ab/CXCR4ab (r=0.116; p=0.506), CXCR5nr/CD49dnr (r=0.001; p=0.996), CXCR5ab/CD49dab (r=0.211; p=0.186), CXCR4nr/CD49dnr (r=0.028; p=0.887) and CXCR4ab/CD49dab (r=0.280; p=0.115). When considering normalized variables, there was no correlation between CXCR5/CXCR4 (r=0.315; p=0.096) and CXCR4/CD49d (r=-0.006; p=0.978), but there was a positive correlation between CXCR5/CD49d (r=0.353; p=0.038). CXCR5 (p=0.707 and 0.343), CXCR4 (p=0.915 and 0.599) and CD49d (p= 0.227 and 0.062) were not significantly associated with MYD88 mutational status, when used the MFI in ab lymphocytes or the value normalized, respectively. There was also no significant association between CXCR5 (p=0.109 and 0.330), CXCR4 (p=0.701 and 0.109) and CD49d (p=0.414 and 0.109) with CXCR4 mutational status, for MFI in ab lymphocytes or the value normalized, respectively.
Conclusion: In our cohort, there was no differences in CXCR5, CXCR4 and CD49d MFI between nr and ab lymphocytes of WM pts. There was a positive correlation between normalized levels of CXCR5 and CD49d, which indicates that an increase in CXCR5 expression is associated with an increase in CD49d expression and suggests a functional relationship between these two markers. There was no significant association between CXCR5, CXCR4 and CD49d with MYD88 and CXCR4 mutational status. The findings propose that behavior of tumor cells may be influenced by adhesion and chemotaxis factors and that malignant cells may use these mechanisms to promote their survival and proliferation, possibly facilitating homing to specific sites in the tumor microenvironment. More studies are needed to validate our findings and to better understand its clinical impact, namely in expansion to secondary lymphoid organs.
Background: Immune cells from rheumatoid arthritis (RA) patients display a reduced in vitro response to Porphyromonas gingivalis (P. gingivalis), which may have functional immune consequences. The aim of this study was to characterize, by flow cytometry, the frequency/activity of monocytes and naturally occurring myeloid dendritic cells (mDCs) in peripheral blood samples from patients with periodontitis and patients with periodontitis and RA. Methods: The relative frequency of monocytes and mDCs in the whole blood, the frequency of these cells producing TNFα or IL-6 and the protein expression levels for each cytokine, before and after stimulation with lipopolysaccharide (LPS) from Escherichia coli plus interferon-γ (IFN-γ), were assessed by flow cytometry, in peripheral blood samples from 10 healthy individuals (HEALTHY), 10 patients with periodontitis (PERIO) and 17 patients with periodontitis and RA (PERIO+RA). Results: The frequency of monocytes and mDCs producing IL-6 or TNF-α and the expression of IL-6 and TNF-α in the PERIO group were generally higher. Within the PERIO+RA group, P. gingivalis and related antibodies were negatively correlated with the monocyte and mDC expression of IL-6. A subgroup of the PERIO+RA patients that displayed statistically significantly lower frequencies of monocytes producing IL-6 after activation presented statistically significantly higher peptidylarginine deiminase (PAD)2/4 activity, anti-arg-gingipain (RgpB) IgG levels, mean probing depth (PD), periodontal inflamed surface area (PISA) and bleeding on probing (BoP). Conclusions: In the patients with PERIO+RA, innate immune cells seemed to produce lower amounts of pro-inflammatory cytokines, which are correlated with worse periodontitis-related clinical and microbiological parameters.
Placenta Accreta Spectrum (PAS) disorders, though uncommon, are a leading cause of maternal morbimortality in modern obstetrics. They can lead to life-threatening complications, such as massive obstetric hemorrhage and hemorrhagic shock, often needing a peripartum hysterectomy. Prenatal diagnosis is crucial for optimal management and outcomes. Still, recent studies show that most cases remain undiagnosed until delivery. Studies suggest that a systematic screening process incorporating prenatal imaging and risk factors can enhance predictive accuracy. This systematic review focuses on better understanding the clinical risk factors that increase the risk for these disorders, thus answering the clinical need for targeted prenatal screening.
Endometrial cancer is one of the most common gynaecological malignancies. Although often diagnosed at an early stage, there is a subset of patients with recurrent and metastatic disease for whom current treatments are not effective. Cancer stem cells (CSCs) play a pivotal role in triggering tumorigenesis, disease progression, recurrence, and metastasis, as high aldehyde dehydrogenase (ALDH) activity is associated with invasiveness and chemotherapy resistance. Therefore, this study aimed to evaluate the effects of ALDH inhibition in endometrial CSCs. ECC-1 and RL95-2 cells were submitted to a sphere-forming protocol to obtain endometrial CSCs. ALDH inhibition was evaluated through ALDH activity and expression, sphere-forming capacity, self-renewal, projection area, and CD133, CD44, CD24, and P53 expression. A mass spectrometry-based proteomic study was performed to determine the proteomic profile of endometrial cancer cells upon N,N-diethylaminobenzaldehyde (DEAB). DEAB reduced ALDH activity and expression, along with a significant decrease in sphere-forming capacity and projection area, with increased CD133 expression. Additionally, DEAB modulated P53 expression. Endometrial cancer cells display a distinct proteomic profile upon DEAB, sharing 75 up-regulated and 30 down-regulated proteins. In conclusion, DEAB inhibits ALDH activity and expression, influencing endometrial CSC phenotype. Furthermore, ALDH18A1, SdhA, and UBAP2L should be explored as novel molecular targets for endometrial cancer.
Introduction
In monoclonal B cell lymphocytosis (MBL) and chronic lymphocytic leukemia (CLL), the expansion of malignant B cells disrupts the normal homeostasis and interactions between B cells and T cells, leading to immune dysregulation. CD20+ T cells are a subpopulation of T cells that appear to be involved in autoimmune diseases and cancer.
Methods
Here, we quantified and phenotypically characterized CD20+ T cells from MBL subjects and CLL patients using flow cytometry and correlated our findings with the B-cell receptor mutational status and other features of the disease.
Results and discussion
CD20+ T cells were more represented within the CD8+ T cell compartment and they showed a predominant memory Tc1 phenotype. CD20+ T cells were less represented in MBL and CLL patients vs healthy controls, particularly among those with unmutated IGVH gene. The expansion of malignant B cells was accompanied by phenotypic and functional changes in CD20+ T cells, including an increase in follicular helper CD4+ CD20+ T cells and CD20+ Tc1 cells, in addition to the expansion of the TCR Vβ 5.1 in CD4+ CD20+ T cells in CLL.
... In solid tumors, such as breast cancer, the diminished responsiveness of NK cells to interferon significantly compromises their function [55]. This is particularly relevant in subtypes such as luminal A, where NK cells activity may be more robust, potentially, influencing treatment outcomes [56]. ...
... Such crosstalk between CLL cells and bystander immune cells might vary among individual patients, leading to abnormal patterns of distribution of normal residual B-and T-cell populations in different tissue compartments, including blood [19][20][21][22][23][24]. In this regard, recent data support a close association between the kinetics of monoclonal B-cell lymphocytosis (MBL) cells and other blood-circulating T-cell populations, such as Tαβ CD4 + CD8 lo and Tαβ CD8 + CD20 + cells [25][26][27][28]. ...
... CSLCs constitute a small subset of tumor cells derived from bulk tumors, possessing stem-like features [6][7][8]. Currently, researchers primarily enrich or identify CSLCs using cell surface markers such as CD133 and CD44 [9], pluripotent transcription factors such as Oct4 and Sox2 [10,11], or sphere formation assays conducted in CSC medium [12][13][14]. It is imperative to thoroughly investigate the specific developmental mechanisms of CSLC, develop new targeted drugs, and uncover novel therapeutic targets. ...
... PB immune cells were identified and characterized by flow cytometry, using a stainlyse-wash procedure previously described [20][21][22]. In summary, 100 µL of PB was incubated with the monoclonal antibodies (mAbs) indicated in Table 4, in the presence of 50 µL of Brilliant stain buffer (Becton Dickinson Biosciences (BD), San Jose, CA, USA), for 30 min in the dark and at room temperature. ...
... GSCs share key properties with normal stem cells, such as self-renewal and differentiation [8,9]. In vitro, GSCs form multicellular three-dimensional (3D) spheroids under non-adherent conditions [10,11]. These 3D spheroids closely mimic solid tumors in vivo, making them valuable for studying GBM biology [12]. ...
... More precisely, gd T cells can harbor different phenotypes, such as: naive, central memory (CM), effector memory (EM) or RA + effector memory (TEMRA) (8,9). Moreover, gd T cells co-express other functional receptors, including activating natural killer receptors (NKR: NKG2D, NKp30 and NKp44) (10,11) and various Toll-like receptors (TLRs) (12). However, they can also express inhibitory NKR such as CD94/NKG2A or immune checkpoints (ICP), such as: PD-1, TIM3, LAG3 or CD39. ...
... First, the antimicrobial activity is attributed to the existence of lysozyme, immunoglobulin, and transferrin in the amniotic fluid 35 . Second, regulation of T-cell function and secretion of an anti-inflammatory agent such as IL-1ra, sTNF, and VEGF-R constitute anti-inflammatory activities of the AM that also exhibit a long-term beneficial effect on the stability of corneal surface and reduce the severity and density of neovascularization 36,37 . Third, AM has shown to be aggressively repopulated by cornea stroma-derived cells, ultimately restoring corneal stromal integrity and helping obviate the necessity of emergent therapeutic keratoplasty in cases with impending or actual corneal perforation 38,39 . ...
... As described in previous studies, 15,27 principal component analysis (PCA) allowed analysis of entire profiles of our assessed osteoclastogenesis-related genes in fibroblasts from each CFU ( Figure 2C and D). Principal components (PCs) with eigenvalues ≥1 were calculated to examine the variations in the fibroblast CFUs as described previously. ...
... One of the most well established effect of SCIT is to induce an initial rise in allergen specific IgE, without significant illness effects, immediately followed by a decline to pre-treatment levels [9][10][11][12]. Furthermore, the amount of receptor bound-IgE in basophils, eosinophils, DCs and monocytes has been shown to be correlated with the levels of serum IgE, due to the FcεRI up-regulation triggered by the binding of IgE to the high affinity receptor [42][43][44][45]. Moreover, SIT is also associated with an increase in IgG levels, particularly allergen-specific IgG1 and IgG4 [14,15]. ...
... 14 Studies have identified an elevated Th17 expression in CSU, while Treg cells may contribute to the autoimmune process. 15,16 Therefore, Th17/Treg immune dysregulation likely plays a crucial role in CSU pathogenesis. However, the correlation between serum Th17/Treg immune dysregulation of CSU patients and the severity of CSU has not been investigated. ...
