Evdoxia Gounari

Cervical cancer remains one of the main causes of female mortality, especially in middle- and low-income countries, despite efforts towards the implementation of global vaccination against human papillomavirus (HPV). The aim of this study was to review and compare the most recently published international guidelines providing recommendations on cervical cancer screening strategies among average and high-risk women. Thus, a comparative review of guidelines by the US Preventive Services Task Force (USPSTF), the American Cancer Society (ACS), the American Society of Clinical Oncology (ASCO), the World Health Organization (WHO), the Canadian Task Force on Preventive Health Care (CTFPHC), the Cancer Council Australia (CCA), and the European Guidelines (EG) was conducted. There is an overall agreement regarding the suggestions made for women younger than 21 and those older than 65, with all guidelines stating against routine screening, with the exceptions of CTFPHC and CCA that expand the age group to up to 70 and 75 years, respectively. Continuation of screening in older women is also suggested in those with a history of a precancerous lesion and those with inadequate screening. Most guidelines recommend routine screening at 30–65 years, while the WHO advises that screening should be prioritized at 30–49 years. HPV DNA testing is the method of choice recommended by most guidelines, followed by cytology as an alternative, except for CTFPHC, which refers to cytology only, with self-sampling being an acceptable method by most medical societies. Agreements exist regarding recommendations for specific groups, such as women with a history of total hysterectomy for benign reasons, women with a complete vaccination against HPV, individuals from the lesbian, gay, bisexual, transgender, and queer communities and women with multiple sexual partners or early initiation of sexual activity. On the other hand, the age group of 21–29 is addressed differently by the reviewed guidelines, while differentiations also occur in the screening strategies in cases of abnormal screening results, in women with immunodeficiency, those with in utero exposure to diethylstilbestrole and pregnant women. The development of consistent practice protocols for the most appropriate cervical cancer screening programs seems to be of major importance to reduce mortality rates and safely guide everyday clinical practice.

Ocrelizumab is a B-cell-depleting monoclonal antibody approved for the treatment of relapsing-remitting multiple sclerosis (RRMS) and active primary progressive MS (aPPMS). This prospective, uncontrolled, open-label, observational study aimed to assess the efficacy of ocrelizumab in patients with aPPMS and to dissect the clinical, radiological and laboratory attributes of treatment response. In total, 22 patients with aPPMS followed for 24 months were included. The primary efficacy outcome was the proportion of patients with optimal response at 24 months, defined as patients free of relapses, free of confirmed disability accumulation (CDA) and free of T1 Gd-enhancing lesions and new/enlarging T2 lesions on the brain and cervical MRI. In total, 14 (63.6%) patients and 13 patients (59.1%) were classified as responders at 12 and 24 months, respectively. Time exhibited a significant effect on mean absolute and normalized gray matter cerebellar volume (F = 4.342, p = 0.23 and F = 4.279, p = 0.024, respectively). Responders at 24 months exhibited reduced peripheral blood ((%) of CD19+ cells) plasmablasts compared to non-responders at the 6-month point estimate (7.69 ± 4.4 vs. 22.66 ± 7.19, respectively, p = 0.043). Response to ocrelizumab was linked to lower total and gray matter cerebellar volume loss over time. Reduced plasmablast depletion was linked for the first time to sub-optimal response to ocrelizumab in aPPMS.

Objectives: Natalizumab (NTZ), a treatment indicated for patients with highly active Relapsing – Remitting Multiple Sclerosis (RRMS), is known to induce increased relative frequency of lymphocytes. Progressive Multifocal Leukoencephalitis (PML) is a rare but serious adverse event related to NTZ. Moreover, reduced L-selectin (CD62L) expression in T-cells in cryopreserved samples of patients with RRMS under NTZ has been proposed as a biomarker of pre-PML state. We explore the association between L-selectin expression in T-cells and hematological parameters in freshly processed samples of patients with RRMS under NTZ. Methods: We studied L-selectin expression in patients with: RRMS under NTZ (n=34), fingolimod (FTY, n=14), interferon-beta (IFNβ, n=22), glatiramer acetate (GA, N=17); in 9 patients with secondary progressive (SP) MS and in 6 healthy controls. Twenty-two patients under NTZ and 6 patients under FTY were followed for 18 months. One NTZ-treated patient developed PML during the study. Results: Patients under NTZ exhibited increased relative frequency of lymphocytes (40.02±1.45) compared to patients under first–line treatment (30.57±1.68, p<0.001) and to patients with SPMS (29±1.56, p=0.02), and a lower mean L-selectin expression in (69.39±1.73) compared to patients under first–line treatment (79.1±1.17, p=0.003). A negative correlation between the relative frequency of CD4+CD62L+ T-cells and the absolute lymphocyte counts (Pearson’s r=0.367, p=0.033) was observed. Discussion: We hereby provide mechanistic insight in a possible pathway implicated in NTZ-related PML risk. These results further underline the need for thorough validation of L-selectin expression in T-cells as a potential pre-PML biomarker.

Peripheral blood (PB) immunophenotyping is commonly required for initial evaluation of various suspected disease entities. Several approaches have been proposed. The objective of this work is to explore the value of a 10color protocol developed in our laboratory for flow cytometric assessment of PB leukocytic subsets, as part of a 2tube screening panel. A combination of CD16/CD56/CD34/CD33/CD19/CD4/CD8/CD3/CD20/CD45 antibodies in 1 tube was applied routinely during flow cytometric analysis of PB samples for diagnostic purposes. The protocol was systematically complemented by a 2nd tube with anti-kappa, anti-lambda, CD5, CD19, and CD45 antibodies for adults and selected pediatric patients, and specifically oriented panels when necessary. 25 samples with no detectable neoplastic PB involvement and 31 samples with a hematolymphoid disorder were investigated retrospectively. The contribution of CD33 in the separation of leukocytic populations, as well as the benefits from the simultaneous assessment of CD20/CD19/CD45, CD16/CD56 and the detection of CD34+ cells were examined. The gating strategy with the use of CD33 provided additional information in certain cases. The protocol enabled recognition of differential expression of CD20 and CD45 in CD19+ cells with chronic lymphocytic leukemia phenotype, overall evaluation of NK and NK like T cells, estimation of CD16− granulocytes and CD56/CD16 expression in monocytes, as well as identification of minor cell subsets, such as CD34+ cells. The proposed 10color combination of antibodies analyzed in a standardized manner can offer significant information in the initial evaluation of PB samples, thus, guiding subsequent investigation if needed.

Background: Plasma cell myeloma (PCM) has been sporadically reported to occur simultaneously or subsequently to mature B lymphoproliferative disorders (LPDs), predominantly chronic lymphocytic leukaemia (CLL). Methods: We describe the clinical and laboratory findings of a 69-year-old male patient who developed plasma cell leukaemia (PCL) 8 years after an initial diagnosis of a low stage CD5+ B LPD and 3 years after treatment for LPD. Results: The transition from a clinically indolent B LPD to an aggressive PCM was documented by bone marrow (BM) biopsy, while flow cytometric (FC) immunophenotyping conferred additional information by disclosing the co-existence of both disorders in BM and the presence of abnormal monotypic PCs in peripheral blood above PCL levels. Phenotypic findings suggested a discrete clonal origin of the two disorders. Conclusions:This report of PCL development in a patient with residual CD5+ B LPD, emphasizes the need for comprehensive diagnostic evaluation of such cases and scrutiny of their aetiological relationship, including FC immunophenotyping due to its high analytical sensitivity and multiparametric capacity compared to morphology or immunohistochemistry alone.

Background: Circulating neoplastic plasma cells (PCs) are a common feature between classical plasma cell myeloma (PCM) and its aggressive variant, plasma cell leukemia (PCL), with their early suspicion and detection being of clinical importance for prognostic and diagnostic purposes.Methods: Morphological and flow cytometric enumeration and characterization of peripheral blood (PB) neoplastic PCs were performed in 3 PCM patients after complete blood count (CBC) evaluation.Results: Early suspicion of circulating PCs by initial CBC scatter plots and monocytosis led to morphological enumeration of PCs, with low level of concordance between morphologists. Flow cytometric PB analysis permitted accurate quantification and characterization of circulating clonal PCs.Conclusions: CBC findings can prompt early suspicion of circulating PCs in PCM patients leading to early and accurate flow cytometric analysis of clonal PCs in addition to morphological PB evaluation. The suggested combined approach proved to be most important in the PCL case with discordant morphological evaluations and marginal PC numbers according to current criteria. © 2016 International Clinical Cytometry Society

In pulmonary sarcoidosis, differential cytokine production in the lungs could be related to variable prognosis of patients at different stages of disease. Twenty patients with pulmonary sarcoidosis (10 at radiographic stage I and 10 at stages II-IV), as well as 10 age-matched healthy volunteers participated in the study. A 4-colour flow cytometric technique was used to measure interferon-γ (IFN-γ), interleukin (IL)-2, tumour necrosis factor-α (TNF-α), IL-4, and IL-13 production in phorbol myristate acetate (PMA)/ionomycin-stimulated CD4+ and CD8+ T cells from bronchoalveolar lavage fluid (BALF) and peripheral blood (PB) of patients, and PB of control subjects. CD4+ T cells from patients showed higher expression of IFN-γ in BALF than in PB. Significant correlations were observed between the percentages of BALF CD4+ and CD8+ T cells expressing intracellular IFN-γ, IL-2, and TNF-α. Stage I patients had lower percentages of IFN-γ-producing CD4+ and CD8+ T cells, as well as TNF-α-producing CD8+ T cells, in BALF (but not in PB) than stage II-IV patients. A decreased TH1 and TC1 response was demonstrated in BALF of patients at stage I of disease, which could explain their anticipated better prognosis.

Several lines of evidence suggest immune system derangement in obstructive sleep apnea syndrome (OSAS) patients. However, no data exist on the long-term effect of continuous positive airway pressure (CPAP) treatment on systemic immunity. Hence, we sought to evaluate this effect on various immunological parameters in OSAS patients. Prospective case series. Sleep unit of a general hospital. Newly-diagnosed, nonsmoking, otherwise healthy OSAS male patients (n = 52) were evaluated on diagnosis and 6 months after CPAP treatment. According to compliance to CPAP use at 6-month follow-up, they were divided into 2 groups: group A (n = 32): good compliance (mean CPAP use ≥ 4 h/night); and group B (n = 20): poor compliance (mean CPAP use < 4 h/night). N/A Blood samples were obtained at baseline and at the 6-month follow-up. Percentage and absolute count of lymphocyte subsets (by flow cytometry), serum TNF-α, IL-6, and uric acid levels were measured. No differences were recorded regarding the baseline anthropometric or sleep characteristics of the 2 groups. In group A, a significant decrease in the absolute count of total lymphocytes (P = 0.003), and of CD4⁺cells (P = 0.001), and a decrease in TNF-α levels (P = 0.001) and uric acid levels (P < 0.001) was observed after CPAP application. On the contrary, no alterations occurred in any of the tested parameters in group B patients. The selective reduction of soluble and cellular immune response factors only in those OSAS patients who exhibited good compliance to CPAP therapy provides further evidence for an ongoing systemic immune process in OSAS.