DataSheet_1_Changes in Treg and Breg cells in a healthy pediatric population.pdf

The interpretation of clinical diagnostic results in suspected inborn errors of immunity, including Tregopathies, is hampered by the lack of age-stratified reference values for regulatory T cells (Treg) in the pediatric population and a consensus on which Treg immunophenotype to use. Regulatory B cells (Breg) are an important component of the regulatory system that have been poorly studied in the pediatric population. We analyzed (1) the correlation between the three immunophenotypic definitions of Treg (CD4+CD25hiCD127low, CD4+CD25hiCD127lowFoxP3+, CD4+CD25hiFoxP3+), and with CD4+CD25hi and (2) the changes in Treg and Breg frequencies and their maturation status with age. We performed peripheral blood immunophenotyping of Treg and Breg (CD19+CD24hiCD38hi) by flow cytometry in 55 healthy pediatric controls. We observed that Treg numbers varied depending on the definition used, and the frequency ranged between 3.3–9.7% for CD4+CD25hiCD127low, 0.07-1.6% for CD4+CD25hiCD127lowFoxP3+, and 0.24-2.83% for CD4+CD25hiFoxP3+. The correlation between the three definitions of Treg was positive for most age ranges, especially between the two intracellular panels and with CD4+CD25hi vs CD4+CD25hiCD127low. Treg and Breg frequencies tended to decline after 7 and 3 years onwards, respectively. Treg’s maturation status increased with age, with a decline of naïve Treg and an increase in memory/effector Treg from age 7 onwards. Memory Breg increased progressively from age 3 onwards. In conclusion, the number of Treg frequencies spans a wide range depending on the immunophenotypic definition used despite a good level of correlation exists between them. The decline in numbers and maturation process with age occurs earlier in Breg than in Treg.</p

Clinical characteristics of patients with recurrent invasive pneumococcal disease.

<p>P, patient identification; PCV-7, 7-valent polysaccharide-protein conjugate vaccine; PCV 13, 13-valent polysaccharide-protein conjugate vaccine; PPV-23, 23-valent pneumococcal polysaccharide vaccine.</p><p>Clinical characteristics of patients with recurrent invasive pneumococcal disease.</p

Microbiological characteristics of patients with recurrent invasive pneumococcal disease.

<p>P, patient identification; ST, sequence type by Multi-locus sequence typing; MDR, Multi-drug resistant strain; PCV-7, 7-valent polysaccharide-protein conjugate vaccine; PCV 13, 13-valent polysaccharide-protein conjugate vaccine; PPV-23, 23-valent pneumococcal polysaccharide vaccine; CSF, cerebrospinal fluid; NA, Not available; MIC, minimal inhibitory concentration; Pen, penicillin; Ery, erythromycin; Cm, chloramphenicol; Tet, tetracycline; Ctx, Cefotaxime; Bold text, resistant strain according to MICs for <i>Streptococcus pneumonia</i>. EUCAST Clinical Meningeal Breakpoints.</p><p>^aNA, ST were not available in episodes identified only by PCR.</p><p>Microbiological characteristics of patients with recurrent invasive pneumococcal disease.</p

Absolute and percentage cell counts of peripheral blood lymphocytes in patient P3.

<p>Absolute and percentage cell counts of peripheral blood lymphocytes in patient P3.</p

Clinical, demographic and genetic characteristics of PMM2-CDG patients.

<p>Clinical, demographic and genetic characteristics of PMM2-CDG patients.</p

Basal numbers of peripheral blood T and NK cells at the time of sample reception.

<p>Basal numbers of peripheral blood T and NK cells at the time of sample reception.</p

Reactivity of NK cell against K562 target cells.

<p>(A) Percentage of degranulated NK cells at 1:1 E/T ratio (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0158863#sec007" target="_blank">Materials and Methods</a> section). (B) Máximal reactivity of NK cells calculated as the maximal number of degranulated CD107a⁺ NK cells (α_max). Values from all tested patients and control subjects are shown. Horizontal lines represent the mean value in each study group. (C) Percentage of degranulated NK cells at 1:1 E/T ratio of fresh and activated NK cells from patient P2 (light grey bars), P5 (white bars) and control subjects (dark bars). Standard deviation in control samples is shown.</p

Level of CD54 expression on different blood lymphocyte subpopulations and monocytes in PMM2-CDG patients and control subjects.

<p>The study was done by flow cytometry. Values are expressed as % MFI <i>vs</i> that observed in controls. CD54 expression was found significantly lower (p = 0.021) in patient B lymphocytes.</p

Primary immunodeficiency associated with chromosomal aberration - an ESID survey

Patients with syndromic features frequently suffer from recurrent respiratory infections, but little is known about the spectrum of immunological abnormalities associated with their underlying chromosomal aberrations outside the well-known examples of Down and DiGeorge syndromes. Therefore, we performed this retrospective, observational survey study.All members of the European Society for Immunodeficiencies (ESID) were invited to participate by reporting their patients with chromosomal aberration (excluding Down and DiGeorge syndromes) in combination with one or more identified immunological abnormalities potentially relating to primary immunodeficiency. An online questionnaire was used to collect the patient data.Forty-six patients were included from 16 centers (24 males, 22 females; median age 10.4 years [range 1.0-69.2 years]; 36 pediatric, 10 adult patients). A variety of chromosomal aberrations associated with immunological abnormalities potentially relating to primary immune deficiency was reported. The most important clinical presentation prompting the immunological evaluation was 'recurrent ear-nose-throat (ENT) and airway infections'. Immunoglobulin isotype and/or IgG-subclass deficiencies were the most prevalent immunological abnormalities reported.Our survey yielded a wide variety of chromosomal aberrations associated with immunological abnormalities potentially relating to primary immunodeficiency. Although respiratory tract infections can often also be ascribed to other causes (e.g. aspiration or structural abnormalities), we show that a significant proportion of patients also have an antibody deficiency requiring specific treatment (e.g. immunoglobulin replacement, antibiotic prophylaxis). Therefore, it is important to perform immunological investigations in patients with chromosomal aberrations and recurrent ENT or airway infections, to identify potential immunodeficiency that can be specifically treated

Additional file 2: of Primary immunodeficiency associated with chromosomal aberration – an ESID survey

Clinical and immunological characteristics of the excluded patients. (DOCX 24 kb