Dariusz Runowski

Th1/Th2 balance and CD45-positive T cell subsets in primary nephrotic syndrome

Abstract T cells are involved in the pathogenesis of nephrotic syndrome (NS). The aim of the study was to determine whether the activity of T-helper-1 (Th1) and T-helper-2 (Th2) cells and the distribution of the lymphocyte subsets, namely CD45RA+CD4+ (”naive” helper T cells, suppressor-inducer), CD45RA+CD8+ (”naive” suppressor T cells, suppressor-effector), CD45RO+CD4+ (”memory” helper T cells), are predictive for steroid sensitivity in children with primary NS. These parameters were assessed at the onset of disease, before initiation of steroid therapy. Two groups of NS children were retrospectively formed according to steroid sensitivity (SS) or resistance (SR). The activity of Th1 and Th2 cells was defined by the production of interleukin-2 (IL-2), interferon-γ, IL-4, and IL-10 in the supernatants of CD4+ T cell cultures activated with autologous monocytes presenting tetanus toxoid (TT). Peripheral lymphocyte subsets were determined using double- or triple-color flow cytometry. In SS children with NS we found a decreased proliferative response of CD4+ T cells to TT stimulation, cytokine synthesis indicating the predominance of Th2 activity, and an increased percentage of activated suppressor-inducer (CD45RA+ CD4+CD25+, 5.18±0.8, P<0.001) and suppressor-effector (CD45RA+CD8+CD25+, 2.05±0.6, P<0.01) cells, with the concomitant reduction of activated memory cells (CD45RO+CD4+CD25+, 0.2±0.1, P<0.001). In children with SRNS we found an increased proliferative response of CD4+ T cells to TT, a rise in activated memory (CD45RO+CD4+CD25+, 3.82±0.7, P<0.01) and suppressor-inducer peripheral T cells (CD45RA+ CD4+CD25+, 3.85±0.6, P<0.01), but a low percentage of activated suppressor-effector (CD45RA+CD8+ CD25+, 0.5±0.2, P<0.05) T cells. We conclude that prior to treatment the distribution of lymphocyte subpopulations in peripheral blood together with Th1 and Th2 cell activity provides a useful tool for evaluating the likelihood of steroid sensitivity in patients with primary NS.

EFFECT OF PSEUDOMONAS AERUGINOSA EXOTOXIN A ON CD3-INDUCED HUMAN T-CELL ACTIVATION

The effect of Pseudomonas aeruginosa (PA) exotoxin A (P-ExA) on CD3-induced T-cell activation was studied on the level of T-cells (proliferation, synthesis of interleukin (IL)-2, expression of IL-2R complex, ICAM-1,2 and LFA-1 molecules), and on the level of monocytes (expression of ICAM-1,2, LFA-1 molecules, as well as FcRI and CD14 receptors). We found that: (1) P-ExA blocked T-cell proliferation and this effect was totally reversed by intact monocytes, and partially by IL-2 or TPA but not by costimulatory cytokines (IL-1alpha, IL-1beta, TNF-alpha or IL-6); (2) P-ExA transiently, in short-term cultures (48 h), inhibited synthesis of IL-2; (3) prolonged stimulation (96 h) of peripheral blood mononuclear cells (PBMC) or CD4 + T-cells with P-ExA in high or low doses (100 and 10 ng/ml, respectively), enhanced the level of IL-2 in the cultures; (4) P-ExA at low dose, combined with IL-1beta, TNF-alpha or IL-6, up-regulated synthesis of IL-2; and (5) stimulation of T-cells with anti-CD3 monoclonal antibody (mAb) and P-ExA at high dose diminished the expression of the p55 chain but not of the p75 chain of IL-2R complex and slightly affected the expression of CD3 complex, ICAM-1,2 and LFA-1 molecules. Hence, P-ExA can regulate the level of IL-2 in cultures of CD3-induced T-cells either by inhibition of IL-2 consumption (when P-ExA is applied in high dose), or by induction of IL-2 production (a costimulatory effect exerted by P-ExA in low dose in combination with monokines). Action of P-ExA on monocytes resulted in: (1) inhibition of the expression of ICAM-1,2 molecules and their ligand LFA-1 molecule; (2) low expression of FcRI receptor (a ligand for Fc part of CD3 mAb); and (3) inhibition (over 90%) of the expression of CD14 molecule. In conclusion, P-ExA-induced anergy of T-cells depends on: (a) decrease in the affinity of IL-2R complex on activated T-cells; and (b) inhibition of the accessory activities of monocytes.

Long-term renal outcome in children with OCRL mutations: retrospective analysis of a large international cohort

Background Lowe syndrome (LS) and Dent-2 disease (DD2) are disorders associated with mutations in the OCRL gene and characterized by progressive chronic kidney disease (CKD). Here, we aimed to investigate the long-term renal outcome and identify potential determinants of CKD and its progression in children with these tubulopathies. Methods Retrospective analyses were conducted of clinical and genetic data in a cohort of 106 boys (LS: 88 and DD2: 18). For genotype-phenotype analysis, we grouped mutations according to their type and localization. To investigate progression of CKD we used survival analysis by Kaplan-Meier method using stage 3 CKD as the end-point. Results Median estimated glomerular filtration rate (eGFR) was lower in the LS group compared with DD2 (58.8 versus 87.4 mL/min/1.73 m2, P < 0.01). CKD stage II-V was found in 82% of patients, of these 58% and 28% had moderate-to-severe CKD in LS and DD2, respectively. Three patients (3%), all with LS, developed stage 5 of CKD. Survival analysis showed that LS was also associated with a faster CKD progression than DD2 (P < 0.01). On multivariate analysis, eGFR was dependent only on age (b = -0.46, P < 0.001). Localization, but not type of mutations, tended to correlate with eGFR. There was also no significant association between presence of nephrocalcinosis, hypercalciuria, proteinuria and number of adverse clinical events and CKD. Conclusions CKD is commonly found in children with OCRL mutations. CKD progression was strongly related to the underlying diagnosis but did not associate with clinical parameters, such as nephrocalcinosis or proteinuria.

Severe acute cardiotoxicity following two intravenous doses of cyclophosphamide in an adolescent treated for rapidly progressive glomerulonephritis

Address for correspondence: Prof. Ryszard Grenda, MD, PhD, The Children’s Memorial Health Institute, Aleja Dzieci Polskich 20, 04–730 Warszawa, Poland, e-mail: r.grenda@ipczd.pl Conflict of interest: none declared Kardiologia Polska Copyright

Ciężkie powikłania i następstwa pierwotnego zespołu nerczycowego u dzieci

Streszczenie Wstep Rokowanie w PZN u dzieci jest dobre pod warunkiem uzyskania i utrzymania remisji. Pogarszają je powiklania, ktore mogą sie zdarzyc w kazdym nawrocie choroby, a takze jako nastepstwa dlugotrwalego bialkomoczu i leczenia. Cel pracy Analiza zagrazających zyciu powiklan i odleglych nastepstw PZN u dzieci. Material i metody U 152 chorych (96 chlopcow, 56 dziewcząt w wieku 0–17,5 roku, średnia 4,10+3,76 r.) hospitalizowanych z powodu ZN w okresie 01.2000–03.2011 badano przebieg kliniczny choroby, obraz patomorfologiczny, wystepowanie ostrych i odleglych powiklan oraz nastepstw PZN. Wyniki U 9 chorych (6,0%; 4 z FSGS, 4 z MCD, 1 z DMP) wystąpilo 12 epizodow OUN, u 2 chorych (1,3%) stwierdzono ChZ-Z, a u 4 (2,7%) – posocznice. PChN z obnizeniem filtracji rozwinela sie u 11 chorych (7,3%; 6 z FSGS, 4 z WZN, 1 z M-PGN), 9 z nich (6,0%) bylo dializowanych, a 7 (4,7%) otrzymalo przeszczep nerki. U 2 z 4 pacjentow z FSGS po przeszczepieniu nastąpil nawrot choroby podstawowej w grafcie. U 2 chorych (1,3%) z DMP stwierdzono chorobe nowotworową: ziarnice zlośliwą u 17-letniej dziewczynki z PSOZN oraz miesaka z komorek poprzecznie prązkowanych jądra u 17-letniego chlopca po 15 latach leczenia SZZN kortykosteroidami, lewamizolem i cyklosporyną A. W czasie obserwacji 4 chorych (2,7%) zmarlo: 2 z powodu posocznicy w przebiegu WZN, 1 – ChZ-Z i 1 – udaru krwotocznego mozgu w okresie leczenia hemodializami. Wnioski Pomimo poprawy rokowania w ZN u dzieci powiklania i nastepstwa odlegle mogą byc groźne dla zycia. Wystepują one niezaleznie od rodzaju glomerulopatii. Leczenie dzieci z ZN powinno odbywac sie pod nadzorem specjalistycznych ośrodkow nefrologicznych dysponujących mozliwościami prowadzenia terapii nerkozastepczej.