Monitoring of Chimerism Following Hematopoietic Stem Cell Transplantation

One of the most important events in the posthematopoietic stem cell transplantation is the immune system reconstitution—a process characterized by a considerable dynamic. During this period, patients are exposed to different life threatening complications. In this chapter, we consider chimerism levels in relation to the conditioning regimens and disease type. Furthermore, the predictive role of chimerism analysis as an important method in monitoring the early diagnosis of graft versus host disease (GVHD), minimal residual disease (MRD), graft failure or rejection, and disease relapse has been discussed

Multiparameter flow cytometric analysis of CD4 and CD8 T cell subsets in young and old people

<p>Abstract</p> <p>Background</p> <p>T cell-mediated immunity in elderly people is compromised in ways reflected in the composition of the peripheral T cell pool. The advent of polychromatic flow cytometry has made analysis of cell subsets feasible in unprecedented detail.</p> <p>Results</p> <p>Here we document shifts in subset distribution within naïve (N), central memory (CM) and effector memory (EM) cells defined by CD45RA and CCR7 expression in the elderly, additionally using the costimulatory receptors CD27 and CD28, as well as the coinhibitory receptors CD57 and KLRG-1, to further dissect these. Although differences between young and old were more marked in CD8 than in CD4 cells, a similar overall pattern prevailed in both. Thus, the use of all these markers together, and inclusion of assays of proliferation and cytokine secretion, may enable the construction of a differentiation scheme applicable to CD4 as well as CD8 cells, with the model (based on Romero et al.) suggesting the progression N→CM→EM1→EM2→pE1→pE2→EM4→EM3→E end-stage non-proliferative effector cells.</p> <p>Conclusion</p> <p>Overall, the results suggest that both differences in subset distribution and differences between subsets are responsible for age-related changes in CD8 cells but that differences within rather than between subsets are more prominent for CD4 cells.</p

Viral load and lymphocyte subpopulations in newly diagnosed patients with chronic Hepatitis B

INTRODUCTION: The immune response against Hepatitis B virus (HBV) represents a key factor for infection outcome. However, the relation between viral replication and host immune reactivity is still a matter of investigation. AIM: To investigate whether the cellular immune response of newly diagnosed treatment naïve chronic hepatitis B (CHB) patients may be influenced by the replicative status of HBV.MATERIALS AND METHODS: A total of 45 (17 female and 28 male) newly diagnosed untreated CHB patients aged 42.48±13.19 years (19÷71 years) were enrolled in this study. The patients were divided in two groups according to the viral load: >0÷≤104 copies/ml (n=25) and >10 4÷<108 copies/ml (n=17). Flowcytometric immunophenotyping was performed for evaluation of the cellular immunity. Serum HBV DNA load was assessed by quantitative real-time polymerase chain reaction.RESULTS: Similar alterations were observed in both patients` groups in comparison with healthy controls which could be summarized as follows: decreased total T cells (CD3+) due to low helper-inducer (CD3+CD4+) and suppressor-cytotoxic (CD3+CD8+) subpopulations; reduced effector cytotoxic (CD8+CD11b-; CD8+CD28+) and activated (CD3+HLA-DR+, CD8+CD38+) T-cell subsets; increased CD57+CD8- cells; elevated percentage of B lymphocytes. No significant differences in the studied immune parameters were detected between both patients` groups except the significantly elevated CD4/CD8 ratio in individuals with higher in comparison to those with lower HBV DNA levels.CONCLUSION: Alterations in the cellular immune repertoire of CHB patients were observed resulting mainly in significantly decreased T-cell subpopulations, particularly those with effector cell immune phenotype regardless of the viral load

Computer Interfaces to Organizations: Perspectives on Borg-Human Interaction Design

We use the term borg to refer to the complex organizations composed of people, machines, and processes with which users frequently interact using computer interfaces and websites. Unlike interfaces to pure machines, we contend that borg-human interaction (BHI) happens in a context combining the anthropomorphization of the interface, conflict with users, and dramatization of the interaction process. We believe this context requires designers to construct the human facet of the borg, a structure encompassing the borg's personality, social behavior, and embodied actions; and the strategies to co-create dramatic narratives with the user. To design the human facet of a borg, different concepts and models are explored and discussed, borrowing ideas from psychology, sociology, and arts. Based on those foundations, we propose six design methodologies to complement traditional computer-human interface design techniques, including play-and-freeze enactment of conflicts and the use of giant puppets as interface prototypes.Comment: 10 page

Care of patients with inborn errors of immunity in thirty J Project countries between 2004 and 2021

IntroductionThe J Project (JP) physician education and clinical research collaboration program was started in 2004 and includes by now 32 countries mostly in Eastern and Central Europe (ECE). Until the end of 2021, 344 inborn errors of immunity (IEI)-focused meetings were organized by the JP to raise awareness and facilitate the diagnosis and treatment of patients with IEI.ResultsIn this study, meeting profiles and major diagnostic and treatment parameters were studied. JP center leaders reported patients’ data from 30 countries representing a total population of 506 567 565. Two countries reported patients from JP centers (Konya, Turkey and Cairo University, Egypt). Diagnostic criteria were based on the 2020 update of classification by the IUIS Expert Committee on IEI. The number of JP meetings increased from 6 per year in 2004 and 2005 to 44 and 63 in 2020 and 2021, respectively. The cumulative number of meetings per country varied from 1 to 59 in various countries reflecting partly but not entirely the population of the respective countries. Altogether, 24,879 patients were reported giving an average prevalence of 4.9. Most of the patients had predominantly antibody deficiency (46,32%) followed by patients with combined immunodeficiencies (14.3%). The percentages of patients with bone marrow failure and phenocopies of IEI were less than 1 each. The number of patients was remarkably higher that those reported to the ESID Registry in 13 countries. Immunoglobulin (IgG) substitution was provided to 7,572 patients (5,693 intravenously) and 1,480 patients received hematopoietic stem cell therapy (HSCT). Searching for basic diagnostic parameters revealed the availability of immunochemistry and flow cytometry in 27 and 28 countries, respectively, and targeted gene sequencing and new generation sequencing was available in 21 and 18 countries. The number of IEI centers and experts in the field were 260 and 690, respectively. We found high correlation between the number of IEI centers and patients treated with intravenous IgG (IVIG) (correlation coefficient, cc, 0,916) and with those who were treated with HSCT (cc, 0,905). Similar correlation was found when the number of experts was compared with those treated with HSCT. However, the number of patients treated with subcutaneous Ig (SCIG) only slightly correlated with the number of experts (cc, 0,489) and no correlation was found between the number of centers and patients on SCIG (cc, 0,174).Conclusions1) this is the first study describing major diagnostic and treatment parameters of IEI care in countries of the JP; 2) the data suggest that the JP had tremendous impact on the development of IEI care in ECE; 3) our data help to define major future targets of JP activity in various countries; 4) we suggest that the number of IEI centers and IEI experts closely correlate to the most important treatment parameters; 5) we propose that specialist education among medical professionals plays pivotal role in increasing levels of diagnostics and adequate care of this vulnerable and still highly neglected patient population; 6) this study also provides the basis for further analysis of more specific aspects of IEI care including genetic diagnostics, disease specific prevalence, newborn screening and professional collaboration in JP countries

Novel multiplex bead-based assay for detection of IDH1 and IDH2 mutations in myeloid malignancies.

Isocitrate dehydrogenase 1 and 2 (IDH) mutations are frequently found in various cancer types such as gliomas, chondrosarcomas and myeloid malignancies. Their molecular detection has recently gained wide recognition in the diagnosis and prognosis of these neoplasms. For that purpose various molecular approaches have been used but a universally accepted method is still lacking. In this study we aimed to develop a novel bead-based liquid assay using Locked nucleic acids (LNA)-modified oligonucleotide probes for multiplexed detection of the most frequent IDH1 (p.R132C, p.R132G, p.R132H, p.R132L, p.R132S) and IDH2 (p.R140Q, p.R172K) mutations. The method includes four steps: 1) PCR amplification of the targeted fragments with biotinylated primers; 2) Direct hybridization to barcoded microbeads with specific LNA-modified oligonucleotide probes; 3) Incubation with phycoerythrin coupled streptavidin; 4) Acquisition of fluorescent intensities of each set of beads on a flow platform (LuminexCorp., USA). We tested the performance of the assay on both artificial plasmid constructs and on clinical samples from 114 patients with known or suspected myeloid malignancies. The method appeared to be superior to direct sequencing having a much higher sensitivity of 2.5% mutant alleles. Applying this method to patients' samples we identified a total of 9 mutations (one IDH1 p.R132C, seven IDH2 p.R140Q and one IDH2 p.R172K). In conclusion, this method could be successfully implemented in the diagnostic work-up for various tumors known to harbor IDH1/2 mutations (e.g. myeloid malignancies, gliomas, etc.). International initiatives are needed to validate the different existing methods for detection of IDH1/2 mutations in clinical settings

Parameters of the fitted linear models shown in Fig. 1.

<p>Parameters of the fitted linear models shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0099769#pone-0099769-g001" target="_blank">Fig. 1</a>.</p

Comparison of the bead-based assay to direct sequencing in selected genomic DNA samples.

<p>The vertical arrows show the positions of the identified mutations.</p

Determination of the sensitivity of the assay as expressed in percent mutant allele.

<p>The horizontal lines denote the threshold level for each mutant's index.</p

Fitted linear standard curves from the test assays for determination of the best performing probe set and the optimal hybridization temperature.

<p>Fitted linear standard curves from the test assays for determination of the best performing probe set and the optimal hybridization temperature.</p