Cytoprotective effect of lithium against spontaneous and induced apoptosis of lymphoid cell line MOLT-4.

Lithium (Li) is still useful in the treatment of bipolar disorder. Cellular mechanisms of Li action are not fully understood and include some cytoprotective properties. Data concerning Li effect on the apoptotic mechanisms in cells other than neurons are fragmentary and contradictory. We have investigated anti-apoptotic activity of Li in a lymphoid derived MOLT-4 cell line. Spontaneous and camptothecin-induced apoptosis was analyzed in cells treated with 0-20 mM Li carbonate. Early apoptosis was identified as significant mitochondrial depolarization (JC-1 staining). Later stages of apoptosis were estimated with annexin V binding and by the proportion of cells containing sub-G1 amounts of DNA (PI staining). We have observed a biphasic effect of Li on the proportion of spontaneously apoptotic cells;namely, low (therapeutic) concentrations of Li had a significant effect stabilizing the mitochondrial membrane polarization, while 10 and 20mM Li increased apoptosis. The latter could be seen both as mitochondrial depolarization as well as an increased proportion of sub-G1 cells, accompanied by reduced proportion of S phase cells. Li at concentrations above 2 mM had a significant, dose-dependent, anti-apoptotic effect on the cells undergoing camptothecin induced apoptosis. In conclusion, demonstrated cytoprotective effect of Li is at least partially related to stabilization of mitochondrial membrane potential and to the reduction of DNA damaging effects in proliferating cells; both may form part of the mechanism through which Li is useful in therapy of bipolar disorder, but may have more general consequences

Hemodialysis Affects Phenotype and Proliferation of CD4-Positive T Lymphocytes

CD4+ T lymphocytes of patients with chronic kidney disease (CKD) are characterized by reduced levels of crucial surface antigens and changes in the cell cycle parameters. Recombinant human erythropoietin (rhEPO) normalizes their altered phenotype and proliferative capacity. Mechanisms leading to the deficient responses of T lymphocytes are still not clear but it is postulated that immunological changes are deepened by hemodialysis (HD). Study of activation parameters of CD4+ T lymphocytes in hemodialyzed and predialysis CKD patients could bring insight into this problem. Two groups of patients, treated conservatively (predialysis, PD) and hemodialyzed (HD), as well as healthy controls, were included into the study; neither had received rhEPO. Proportions of main CD4+CD28+, CD4+CD25+, CD4+CD69+, CD4+CD95+, and CD4+HLA-DR+ lymphocyte subpopulations and proliferation kinetic parameters were measured with flow cytometry, both ex vivo and in vitro. No differences were seen in the proportions of main CD4+ lymphocyte subpopulations (CD4+CD28+, CD4+CD25+, CD4+HLA-DR+, CD4+CD69+, CD4+CD95+) between all examined groups ex vivo. CD4+ T lymphocytes of HD patients exhibited significantly decreased expression of co-stimulatory molecule CD28 and activation markers CD25 and CD69 after stimulation in vitro when compared with PD patients and healthy controls. HD patients showed also decreased percentage of CD4+CD28+ lymphocytes proliferating in vitro; these cells presented decreased numbers of finished divisions after 72 h of stimulation in vitro and had longer G0→G1 time when compared to healthy controls. CD4+ T lymphocytes of PD patients and healthy controls were characterized by similar cell cycle parameters. Our study shows that repeated hemodialysis procedure influences phenotype and proliferation parameters of CD4+ T lymphocytes

Absence of the suprarenal segment of the inferior vena cava with a coexisting absence of the right brachiocephalic vein in a 22-year-old Caucasian male with arterial hypertension

Congenital anomalies of the inferior vena cava (IVC) are rarely observed malformations of the venous system, occurring in 0.3% of otherwise healthy individuals, and in 0.6% to 2% of patients with coexisting cardiovascular defects. They are usually asymptomatic and recognised incidentally during imaging, operations or dissection studies. In this paper we report an extremely rare case of a 22-year-old Caucasian male, admitted for the purpose of excluding secondary causes of hypertension. During imaging of the abdomen and the thorax we found a complete lack of the suprarenal segment of the IVC, with a coexisting absence of the right brachiocephalic vein. We discuss the problem of congenital defects of the IVC and we review the relevant literature

Different distribution of CD4 and CD8 T cells in synovial membrane and peripheral blood of rheumatoid arthritis and osteoarthritis patients.

Rheumatoid arthritis (RA) and osteoarthritis (OA) are chronic diseases associated with morphological joint changes. Synovial membrane (SM) involvement was established for RA, but the data for OA are limited, because OA is usually regarded as noninflammatory disease. Changes in immune system in RA are not limited to joints, and the significant role of T cells of peripheral blood (PB) is not disputable. However, there is still an open debate about PB immunological profile in OA. Therefore, we decided to measure the distribution of CD4+ and CD8+ T cells, regarding CD28 expression, both in PB and SM of RA and OA patients, on the same day. Altogether, eleven RA patients, 11 OA patients and similar numbers of age-matched healthy controls were included into the study. Flow cytometry was used for T cells subpopulation distinguishing and quantification; monoclonal antibodies against CD3, CD4, CD8 and CD28 with different fluorochromes were used for stainings. The RA patients had significantly higher percentage of CD3+4+ cells in PB as compared to OA patients and relevant control group. Both within the CD4+ and CD8+ compartments, significantly lower percentages of cells bearing the CD28 marker were found in the PB of OA as compared to RA patients. The proportion of CD3+CD4+ cells in SM was dependent on age of OA patients, older OA patients had significantly higher value of their SM/blood ratio than RA patients. Older OA subjects were also characterized by higher values of the SM/blood ratio of both CD4+CD28+ and CD8+CD28+ subpopulations than RA or younger OA patients. In conclusion, in contrast to the traditional view of OA disease, our results give support to the hypothesis that OA may also (like RA) be a disease with a local immunological involvement

Wpływ rekombinowanej ludzkiej erytropoetyny na układ odpornościowy

Erytropoetyna (EPO) jest hormonem produkowanym przez komórki śródmiąższowe nerki, który działa na komórki macierzyste erytrocytów, regulując ich proliferację i chroniąc przed apoptozą. Ze względu na budowę upodobniającą ją do czynników tkankowych o charakterze cytokin uważa się, że EPO poza wpływem na układ czerwonokrwinkowy może też regulować pracę układu odpornościowego. Przemawiają za tym wyniki licznych badań przeprowadzonych wśród chorych na przewlekłą chorobę nerek (PChN), którym podaje się rekombinowaną ludzką erytropoetynę (rhEPO) w celu wyrównania niedokrwistości, która jest nieodłącznym elementem tej choroby. W publikacji omówiono zaburzenia funkcjonowania układu odpornościowego z uwzględnieniem różnych rodzajów odpowiedzi immunologicznej. Przedstawiono też wyniki badań dotyczących pozytywnego wpływu leczenia za pomocą rhEPO u chorych hemodializowanych, które wskazują na istnienie wielu mechanizmów poprawy odpowiedzi immunologicznej u tych pacjentów. Artykuł ma na celu zwrócić uwagę na fakt, że poprawa funkcjonowania układu odpornościowego u pacjentów z PChN nie wydaje się być jedynie efektem korekcji niedokrwistości. Przemawia za tym zmiana ekspresji antygenów na limfocytach, dzięki którym możliwe są ich wzajemne relacje, poprawa proliferacji limfocytów T oraz zmiany w profilu cytokin u tych chorych. Owe zmiany mogą wynikać z naśladowania przez EPO działania innych cytokin bądź bezpośredniego wpływu na limfocyty i monocyty poprzez receptor dla EPO lub inne receptory z rodziny receptorów hematopoetyn, które charakteryzują się podobną budową i przekazują sygnał przez podobny system sygnalizacyjny w komórce

Influence of oxygen concentration on T cell proliferation and susceptibility to apoptosis in healthy men and women

Introduction. Much of what we know about the functioning of human T lymphocytes is based on the experiments carried out in atmospheric oxygen (O2) concentrations, which are significantly higher than those maintained in blood. Interestingly, the gender differences in the activity of T cells and their susceptibility to apoptosis under different O2 conditions have not yet been described. The aim of the study was to compare two main markers of lymphocyte function: proliferation capacity and ability to produce cytokines as well as their susceptibility to apoptosis under two different O2 concentrations, between men and women. Materials and methods. 25 healthy volunteers, both males (13) and females (12) were recruited to the study (mean age 25.48 ± 5.51). By using cytometry proliferation parameters of human CD4+ CD28+ cells or CD8+CD28+ cells in response to polyclonal stimulation of the TCR/CD3 complex at atmospheric (21%) and physiological (10%) O2 concentrations using our modified dividing cell tracking technique (DCT) were analyzed as well as the percentages of apoptotic cells. We also determined the levels of IFN-γ, IL-2, IL-10 and IL-17A using Cytometric Bead Array Flex system in cell culture supernatants. Results. CD4+CD28+ and CD8+CD28+ cells from the whole study group were characterized by shorter time required to enter the first (G1) phase of the first cell cycle at 21% compared to 10% O2. Both T cell populations performed significantly more divisions at 21% O2. The percentages of dividing cells were also significantly higher at atmospheric O2. Interestingly, data analysis by gender showed that male lymphocytes had similar proliferative parameters at both O2 concentrations while female lymphocytes proliferate more efficiently (note from the author: we cannot say that lymphocytes proliferate faster, rather more effectively, because cells perform more divisions, which gives more percentage of offspring cells) at 21% oxygen. Compared to males, the female CD4+ cells showed increased susceptibility to apoptosis at both O2 concentrations. No differences in the levels of cytokines regardless of gender and oxygen conditions were found. Conclusions. We showed that in vitro female T cells (both CD4+ and CD8+ cells) are more sensitive than male lymphocytes to low O2 concentration as demonstrated by the decrease in their proliferation dynamics. The effect does not depend on increased apoptosis of female T cells under low O2 because percentage of apoptotic cells was similar at both O2 concentrations

Expression of calpain-calpastatin system (CCS) member proteins in human lymphocytes of young and elderly individuals; pilot baseline data for the CALPACENT project.

Ubiquitous system of regulatory, calcium-dependent, cytoplasmic proteases – calpains – and their endogenous inhibitor – calpastatin – is implicated in the proteolytic regulation of activation, proliferation, and apoptosis of many cell types. However, it has not been thoroughly studied in resting and activated human lymphocytes yet, especially in relation to the subjects’ ageing process. The CALPACENT project is an international (Polish-Italian) project aiming at verifying the hypothesis of the role of calpains in the function of peripheral blood immune cells of Polish (Pomeranian) and Italian (Sicilian) centenarians, apparently relatively preserved in comparison to the general elderly population. In this preliminary report we aimed at establishing and comparing the baseline levels of expression of μ- and m-calpain and calpastatin in various, phenotypically defined, populations of human peripheral blood lymphocytes for healthy elderly Sicilians and Poles, as compared to these values observed in young cohort

Rola limfocytów B w mechanizmach patogenezy wybranych chorób

Układ immunologiczny zdrowego człowieka ma za zadanie obronę oraz utrzymywanie homeostazy organizmu poprzez kontrolowanie antygenów zarówno wnikających do ustroju, jak i tych, które są w nim produkowane. Do najważniejszych komórek pełniących taką funkcję należą ściśle ze sobą współistniejące różne populacje limfocytów. To od nich zależą rozpoznanie antygenu i reakcja mająca na celu jego neutralizację. Jedną z form tej aktywności neutralizującej potencjalny patogen jest produkcja przeciwciał, których źródłem są limfocyty B. Zaburzenia funkcji tych tak zwanych efektorów odpowiedzi humoralnej (prowadzące w konsekwencji do zaburzeń liczby i swoistości przeciwciał) są podstawą (elementem patogenezy) wielu chorób i jako takie stały się celem licznych prac badawczych. Liczebność i proporcje klas limfocytów B, wielkość ich odpowiedzi na stymulację oraz poziom, klasy i swoistość wytwarzanych przeciwciał są bardzo ważnymi czynnikami diagnostycznymi i prognostycznymi w tak na pozór różnych chorobach, jak między innymi: przewlekła obturacyjna choroba płuc, astma oskrzelowa, śródmiąższowe zapalenie płuc, rak płuc czy, co oczywiste, liczne choroby autoimmunologiczne. Fakt ten podkreśla uniwersalność zastosowania obserwacji limfocytów B w kontroli przebiegu wielu schorzeń i ponownie wykazuje, że stan układu immunologicznego odzwierciedla stan zdrowia danej osoby. Forum Medycyny Rodzinnej 2011, tom 5, nr 1, 37-4

The practice of tredmii-test using to teenagers with arterial hypertensia and different body weight

The purpose. Definition of influence of physical activities on arterial pressure and physical working capacity at teenagers with various weight of a body. Materials and methods. 118 teenagers with various degree of adiposity and an arterial hypertensia have been subjected to the tredmil-test. Raw data and physical activity response are defined. According to the results of this test direct and indirect indexes defining physical efficiency are calculated. The comparison of indexes depending on the adiposity degree is carried out. The conclusion.The direct dependence of body weight and arterial pressure and the inverse relationship of body weight and physical efficiency are revealed.Цель. Определение влияния физических нагрузок на артериальное давление и физическую работоспособность у подростков с различной массой тела. Материалы и методы. Проведена тредмилметрия у 118 подростков с различной степенью ожирения и артериальной гипертензией. Определены исходные данные и реакция на физическую нагрузку. По результатам проведенного теста вычислены прямые и косвенные показатели, определяющие физическую работоспособность у подростков в зависимости от степени ожирения. Заключение. Выявлена прямая зависимость массы тела и величин артериального давления и обратная зависимость массы тела и физической работоспособности