Stężenie interleukiny 2 i interleukiny 10 u pacjentów z cukrzycą typu 2 i rakiem okrężnicy

Introduction. The risk of colon cancer (CC) develop­ment is increased significantly among patients with type 2 diabetes (T2DM). A mechanism responsible for a higher prevalence of CC among diabetic patients may be associated with disturbances of the immune system. Cytokines — interleukin-2 (IL-2) and interleukin-10 (IL- -10) play relevant role in the immune response. The aim of this study was to investigate the differences in the immunological state in terms of IL-2 and IL-10 levels among groups of patients with T2DM, patients with CC, patients with T2DM and CC and patients without these diseases. Material and methods. 80 patients were included in the tests and split into 4 groups: group 1 — 24 patients with T2DM, group 2 — 24 patients with CC, group 3 — 10 patients with CC and T2DM, and group 4 — 22 persons without T2DM or CC. Colonoscopy was per­formed for all the patients. All cases of colon cancer were confirmed by histopathological examination. Laboratory measurements included blood tests such as fasting glucose, insulin, C-peptide and HbA1c. The serum concentration of IL-2 and IL-10 was determined by the immunoenzymatic (ELISA) method. Results. The concentration of IL-2 was statistically higher in the group of patients with T2DM and CC than in the groups of patients without those diseases (4.21 ± 1.61 SE pg/ml vs. group 1 — 1.57 ± 0.44 SE pg/ml, group 2 — 1.64 ± 0.27 SE pg/ml, group 4 — 1.95 ± 0.47 SE pg/ml; p &lt; 0.05). There were no statistically significant differences in the concentrations of IL-10 in patients with T2DM and CC compared with other subjects. The level of fasting glucose and HbA1c in the groups of patients with T2DM (group 1) and T2DM with CC (group 3) was statistically higher than in the groups of patients without T2DM. There were no statistically significant differences between the groups in levels of insulin, C-peptide and HOMA-IR. Conclusions. The concentration of IL-2 was statistically higher in the group of patients with T2DM and colon cancer than in other groups. Elevated level of IL-2 can be a marker of an increased risk of CC in people with type 2 diabetes. It might be useful in indicating a group of patients with differences in immune system particu­larly susceptible to the development of colon cancer.  Wstęp. U chorych na cukrzycę stwierdza się istotnie zwięk­szone ryzyko raka jelita grubego. Mogą być za to odpo­wiedzialne m.in. zaburzenia układu immunologicznego. Istotną rolę w odpowiedzi immunologicznej odgrywają cytokiny — interleukina 2 (IL-2) i interleukina 10 (IL-10). Celem pracy była ocena stężenia we krwi wybranych cy­tokin — IL-2 oraz IL-10 — u pacjentów z cukrzycą typu 2 i rakiem jelita grubego w porównaniu z osobami z cukrzy­cą typu 2 bez raka jelita grubego, z rakiem jelita grubego bez cukrzycy oraz bez obu tych patologii. Materiał i metody. Do badania włączono 80 chorych, których podzielono na 4 grupy — grupa 1 (24 osoby) z cukrzycą typu 2, grupa 2 (24 osoby) — z rakiem jelita grubego bez cukrzycy, grupa 3 (10 osób) —– z rakiem jelita grubego oraz cukrzycą typu 2, grupa 4 (22 oso­by) — grupa kontrolna bez cukrzycy oraz bez raka jelita grubego. Wszyscy pacjenci mieli wykonaną kolonoskopię. U osób z nowotworem jelita grubego potwierdzono rozpo­znanie w badaniu histopatologicznym. Przeprowadzono badania laboratoryjne obejmujące ocenę glikemii na czczo, stężenie insuliny i peptydu C oraz odsetek hemoglobiny glikowanej HbA1c. Stężenie IL-2 i IL-10 oznaczano metodą immunoenzymatyczną, stosując zestawy Human IL-2 ELISA KIT i Human IL-10 (DIACLONE Research, Francja). Wyniki. W grupie osób z cukrzycą typu 2 i rakiem jelita grubego stwierdzono statystycznie wyższe wartości stężenia interleukiny 2 (4,21 ± 1,61 SE pg/ml) w porów­naniu z innymi grupami pacjentów (grupa 1 — 1,57 ± 0,44 SE pg/ml, grupa 2 — 1,64 ± 0,27 SE pg/ml, grupa 4 — 1,95 ± 0,47 SE pg/ml; p < 0,05). Nie stwierdzono istotnej statystycznie różnicy pomiędzy badanymi grupami pod względem stężeń IL-10. Obserwowano statystycznie istotny wyższy poziom glikemii na czczo oraz hemoglobiny glikowanej w grupach z cukrzycą typu 2 oraz z cukrzycą typu 2 z towarzyszącym rakiem jelita grubego. Nie wykazano statystycznie istotnej różnicy pomiędzy grupami w poziomach insulinemii, peptydu C ani wskaźnika HOMA-IR. Wnioski. W grupie osób z cukrzycą typu 2 i rakiem jelita grubego stwierdzono statystycznie wyższe war­tości stężenia interleukiny 2 w porównaniu z innymi grupami. Podwyższone stężenie IL-2 w surowicy krwi może być wskaźnikiem zwiększonego ryzyka raka jelita grubego u osób z cukrzycą typu 2. Można zasugero­wać, że pacjenci z cukrzycą typu 2 oraz podwyższonym stężeniem IL-2 w surowicy krwi powinni być objęci szczególnym nadzorem onkologicznym

Decreased Production of TNF-α and IL-6 Inflammatory Cytokines in Non-Pregnant Idiopathic RPL Women Immunomodulatory Effect of Sildenafil Citrate on the Cellular Response of Idiopathic RPL Women

Sildenafil citrate (SC), a PDE5 inhibitor, a drug for erectile dysfunction (ED) and pulmonary hypertension (PAH), was found to exert a positive effect on pregnancy outcomes when administered intravaginally before conception. In our previous studies, sildenafil increased endometrial thickness and significantly decreased peripheral blood NK cell activity after the intravaginal administration in women with recurrent pregnancy loss (RPL). No data are available to confirm the effect of sildenafil on maternal T cell populations involved in shaping fetal-maternal tolerance and NK cell activity. Thus, the present study aimed to establish if SC influences NKT cells or the axis of Th17/Treg cells and Th1/Th2 cytokine production. Materials and methods: Twenty-one healthy fertile women and twenty-two nonpregnant women with idiopathic RPL were studied. The ELISA method was used to evaluate the production of cytokines, including IL-2, IL-12p40, IL-4, IL-10, IL-6, IL-17, IL-21, TGF-β, TNF-α, and IFN-γ in PBMC culture supernatants before and after supplementation with the physiological concentration of SC. The percentages of NKT (CD56+CD3+CD44+CD161+), Treg (CD4+CD25+FOXP3+) and Th17 (CD4+CD25+IL-17A+) cells were determined with flow cytometry method. Results: Unexpectedly, we found that the PBMCs of patients with RPL produced a significantly lower level of inflammatory cytokines (TNF-α and IL-6) and a higher level of anti-inflammatory cytokines (TGF-β and IL-10). SC significantly decreased IL-6, IL-12 and increased TGF-β cytokine concentration in fertile women. In the case of RPL patients’ PBMCs, SC improved the production of TNF-α and IL-10. Conclusions: Lower concentration of proinflammatory cytokines in idiopathic RPL women compared to fertile women might suggest the exhaustion of the immune system. The emphasized production of IL-10 by SC partially explains the previously observed downregulation of NK cell activity in RPL patients. The immunomodulatory effect of the drug might be utilized in anti-inflammatory therapies and help achieve positive pregnancy outcomes in women with reproductive failure due to a Th1/Th2 imbalance

Sildenafil Can Affect Innate and Adaptive Immune System in Both Experimental Animals and Patients

Sildenafil, a type 5 phosphodiesterase inhibitor (PDE5-I), is primarily used for treating erectile dysfunction. Sildenafil inhibits the degradation of cyclic guanosine monophosphate (cGMP) by competing with cGMP for binding site of PDE5. cGMP is a secondary messenger activating protein kinases and a common regulator of ion channel conductance, glycogenolysis, and cellular apoptosis. PDE5 inhibitors (PDE-Is) found application in cardiology, nephrology, urology, dermatology, oncology, and gynecology. Positive result of sildenafil treatment is closely connected with its immunomodulatory effects. Sildenafil influences angiogenesis, platelet activation, proliferation of regulatory T cells, and production of proinflammatory cytokines and autoantibodies. Sildenafil action in humans and animals appears to be different. Surprisingly, it also acts differently in males and females organisms. Although the immunomodulatory effects of PDE5 inhibitors appear to be promising, none of them reached the point of being tested in clinical trials. Data on the influence of selective PDE5-Is on the human immune system are limited. The main objective of this review is to discuss the immunomodulatory effects of sildenafil in both patients and experimental animals. This is the first review of the current state of knowledge about the effects of sildenafil on the immune system

Differences in the Expression of KIR, ILT Inhibitory Receptors, and VEGF Production in the Induced Decidual NK Cell Cultures of Fertile and RPL Women

Problem. Natural killer (NK) cells are the most abundant leukocyte population in the uterus. The interactions of the maternal NK expression of killer cell immunoglobulin-like receptors (KIRs) and human inhibitory receptor Ig-like transcript (ILT) with fetal HLA determine the activation of NK cells and pregnancy outcomes. Moreover, dNK cells release numerous angiogenic factors including VEGF. Our previous study showed that sildenafil citrate (SC) significantly decreased peripheral blood NK (pbNK) cell activity and improved intrauterine blood flow, which correlated with a successful pregnancy outcome. Thus, in this study, we investigated whether SC influenced the expression of KIR or ILT receptors on induced decidual NK (idNK), the apoptosis of cells, and VEGF-A production in the culture supernatants of idNK cells. Method of Study. pbNK cells from 24 healthy women and 23 women with RPL were converted to idNK cells under hypoxia, IL-15, and TGF-β conditions. The cultures were prepared with or without SC. Changes in KIR2DL1 (CD158a), NKG2A (CD159a), ILT-2 (CD85j), and ILT-4 (CD85d) expression on CD56+CD16- cells and their apoptosis were determined via flow cytometry. VEGF-A level was established in culture supernatants with the ELISA method. Results. KIR2DL1 and ILT-2 expression on idNK cells was higher in healthy women than in RPL patients. Sildenafil enhanced NKG2A expression in RPL patients. VEGF concentration was higher in fertile woman idNK cell cultures. idNK cells were more sensitive for necrosis in RPL than in fertile women. SC did not influence VEGF production or idNK cell apoptosis. Conclusions. A combination of hypoxia, IL-15, and AZA promotes the conversion of pbNK into idNK cells CD56+CD16--expressing KIR receptors and produces VEGF. Alterations in KIR2DL1 and ILT-2 expression as well as impaired VEGF production were associated with RPL. SC affects NKG2A expression on RPL idNK cells. SC had no effect on VEGF release or idNK cell apoptosis

Differences in Immune Checkpoints Expression (TIM-3 and PD-1) on T Cells in Women with Recurrent Miscarriages—Preliminary Studies

Background: Immune checkpoints are molecules that regulate the function of immune cells and control inflammation processes. An important role in this regard is played by TIM-3/Gal-9 and PD-1/PDL-1 interactions. Previous research performed in a mouse model of pregnancy loss confirmed that blocking TIM-3 could induce fetal loss. Similarly, the PD-1 molecule maintains protective interactions between the mother’s immune cells and the fetus. The purpose of this study was to assess the expression of these molecules on a range of T lymphocyte subpopulations from non-pregnant women with recurrent spontaneous abortion (RSA) versus healthy fertile women. Methods: PBMCs were isolated by gradient centrifugation of blood obtained from 12 healthy women and 24 women with RSA and immediately stained for flow cytometry analysis. Standard immunophenotyping of PBMC was performed with the antibodies against classical lymphocyte markers: CD3, CD4, CD8, and CD56. Immune checkpoints were investigated using antibodies against PD-1(CD279) and TIM-3(CD366). Results: We found that expression of TIM-3 was significantly decreased on CD8+ T lymphocytes in the RSA group, and expression of PD-1 was upregulated on CD4+ T lymphocytes in the RSA group in comparison to the healthy controls. Conclusions: Considering our findings, therapeutic intervention towards immune checkpoints may be a promising treatment option for recurrent spontaneous abortion

Sildenafil Citrate Downregulates PDE5A mRNA Expression in Women with Recurrent Pregnancy Loss without Altering Angiogenic Factors—A Preliminary Study

In our previous study, we showed that sildenafil citrate (SC), a selective PDE5A blocker, modulated NK cell activity in patients with recurrent pregnancy loss, which correlated with positive pregnancy outcomes. It was found that NK cells had a pivotal role in decidualization, angiogenesis, spiral artery remodeling, and the regulation of trophoblast invasion. Thus, in the current study, we determined the effects of SC on angiogenic factor expression and production, as well as idNK cell activity in the presence of nitric synthase blocker L-NMMA. Methods: NK cells (CD56+) were isolated from the peripheral blood of 15 patients and 15 fertile women on MACS columns and cultured in transformation media containing IL-15, TGF-β, and AZA—a methylation agent—for 7 days in hypoxia (94% N2, 1% O2, 5% CO2). Cultures were set up in four variants: (1) with SC, (2) without SC, (3) with NO, a synthase blocker, and (4) with SC and NO synthase blocker. NK cell activity was determined after 7 days of culturing as CD107a expression after an additional 4h of stimulation with K562 erythroleukemia cells. The expression of the PDE5A, VEGF-A, PIGF, IL-8, and RENBP genes was determined with quantitative real-time PCR (qRT-PCR) using TaqMan probes and ELISA was used to measure the concentrations of VEGF-A, PLGF, IL-8, Ang-I, Ang-II, IFN–γ proteins in culture supernatants after SC supplementation. Results: SC downregulated PDE5A expression and had no effect on other studied angiogenic factors. VEGF-A expression was increased in RPL patients compared with fertile women. Similarly, VEGF production was enhanced in RPL patients’ supernatants and SC increased the concentration of PIGF in culture supernatants. SC did not affect the expression or concentration of other studied factors, nor idNK cell activity, regardless of NO synthase blockade

Levels of interleukin-2 in patients with colon cancer and diabetes type 2

Introduction and objective: The risk of development colon cancer (CC) is increased significantly among patients with the type 2 diabetes (T2DM). A mechanism responsible for the higher prevalence of CC among diabetic patients may be associated with the immunity system. The aim of this study is to point out the differences in the immunity state in terms of interleukin 2 level among patients with T2DM suffering from CC, and patients without these diseases Material and Methods: 79 patients were included the tests, divided into 4 groups: Group 1–23 people with T2DM, Group 2–23 people with large intestine CC, Group 3–10 people with large CC and T2DM, and Group 4–23 people without T2DM or CC. Each patient had a colonoscopy and those with cancer were confirmed in a histopathological examination. Laboratory measurements included fasting glucose, insulin, C-peptide. The concentration of interleukin-2 in serum was determined with the immunoenzymatic (ELISA) method. Results: The results obtained showed that in patients with T2DM and CC the concentration of interleukin-2 was statistically higher than in the other groups.(4.21±1.61 pg/ml vs. Group 1 -1.64±0.44 pg/ml, Group 2–1.54±0.21 pg/ml, and Group 4–1.70±0.36 pg/ml; p<0.05). Insulin levels, C-peptide and HOMA-IR did not differ significantly between groups, but a tendency was observed to higher values of HOMA-IR and insulin levels in the groups with T2DM alone and T2DM with concomitant CC. Conclusions: The data show differences in the immunity state of patients with T2DM and CC, compared with people without those two diseases. Elevated level of interleukin 2 found in this group, after confirmation in other studies with more patients, could be used as a marker of an increased risk of CC in people with T2D

Poziom interleukiny-2 i interleukiny-10 u pacjentów z cukrzycą typu 2 i rakiem okrężnicy.

  Introduction. The risk of colon cancer (CC) develop­ment is increased significantly among patients with type 2 diabetes (T2DM). A mechanism responsible for a higher prevalence of CC among diabetic patients may be associated with disturbances of the immune system. Cytokines — interleukin-2 (IL-2) and interleukin-10 (IL- -10) play relevant role in the immune response. The aim of this study was to investigate the differences in the immunological state in terms of IL-2 and IL-10 levels among groups of patients with T2DM, patients with CC, patients with T2DM and CC and patients without these diseases. Material and methods. 80 patients were included in the tests and split into 4 groups: group 1 — 24 patients with T2DM, group 2 — 24 patients with CC, group 3 — 10 patients with CC and T2DM, and group 4 — 22 persons without T2DM or CC. Colonoscopy was per­formed for all the patients. All cases of colon cancer were confirmed by histopathological examination. Laboratory measurements included blood tests such as fasting glucose, insulin, C-peptide and HbA1c. The serum concentration of IL-2 and IL-10 was determined by the immunoenzymatic (ELISA) method. Results. The concentration of IL-2 was statistically higher in the group of patients with T2DM and CC than in the groups of patients without those diseases (4.21 ± 1.61 SE pg/ml vs. group 1 — 1.57 ± 0.44 SE pg/ /ml, group 2 — 1.64 ± 0.27 SE pg/ml, group 4 — 1.95 ± 0.47 SE pg/ml; p &lt; 0.05). There were no statistically significant differences in the concentrations of IL-10 in patients with T2DM and CC compared with other subjects. The level of fasting glucose and HbA1c in the groups of patients with T2DM (group 1) and T2DM with CC (group 3) was statistically higher than in the groups of patients without T2DM. There were no statistically significant differences between the groups in levels of insulin, C-peptide and HOMA-IR. Conclusions. The concentration of IL-2 was statistically higher in the group of patients with T2DM and colon cancer than in other groups. Elevated level of IL-2 can be a marker of an increased risk of CC in people with type 2 diabetes. It might be useful in indicating a group of patients with differences in immune system particularly susceptible to the development of colon cancer.  Przesłanki i cele pracy U chorych na cukrzycę stwierdza się istotnie zwiększone ryzyko raka jelita grubego. Mogą być za to odpowiedzialne m.in. zaburzenia układu immunologicznego. Istotną rolę w odpowiedzi immunologicznej odgrywają cytokiny - interleukina 2 (IL-2), interleukina 10 (IL-10), Celem pracy była ocena stężenia we krwi wybranych cytokin - interleukiny 2, interleukiny 6, interleukiny 10 u pacjentów z cukrzycą typu 2 i rakiem jelita grubego w porównaniu z osobami z cukrzycą typu 2 bez raka jelita grubego, z rakiem jelita grubego bez cukrzycy oraz bez obu tych patologii. Materiał i metody Do badania włączono 80 chorych, których podzielono na 4 grupy- 1 grupa (24 osoby) z cukrzycą typu 2, 2 grupa (24 osoby)- z rakiem jelita grubego bez cukrzycy, 3 grupa (10 osób) – z rakiem jelita grubego oraz cukrzycą typu 2, 4 grupa (22 osoby)– kontrolna bez cukrzycy oraz bez raka jelita grubego. Wszyscy pacjenci mieli wykonaną kolonoskopię, osoby z nowotworem jelita grubego miały potwierdzenie rozpoznania w badaniu histopatologicznym. Były wykonane badania laboratoryjne obejmujące ocenę glikemii na czczo, stężenie insuliny, peptydu C, hemoglobiny glikowanej HbA1c. Stężenie interleukiny i interleukiny 10 było mierzone metodą immunoenzyatyczną Human IL-2 ELISA KIT, Human IL-10 (DIACLONE Research, France). Wyniki W grupie osób z cukrzycą typu 2 i rakiem jelita grubego stwierdzono statystycznie wyższe wartości stężenia interleukiny-2 (4.21±1.61 SE pg/ml) w porównaniu z innymi grupami pacjentów (grupa 1 - 1.57±0.44 SE pg/ml, grupa 2 - 1.64±0.27 SE pg/ml, grupa 4 – 1.95±0.47 SE pg/ml). Nie stwierdzono istotnej statystycznie różnicy stężeń IL-10 pomiędzy badanymi grupami. Obserwowano statystycznie istotny wyższy poziom glikemii na czczo oraz hemoglobiny glikowanej w grupach z cukrzycą typu 2 oraz z cukrzycą typu 2 z towarzyszącym rakiem jelita grubego. Nie wykazano statystycznie istotnej różnicy pomiędzy grupami w poziomach insulinemii, peptydu C czy wskaźnika HOMA-IR. Wnioski • W grupie osób z cukrzycą typu 2 i rakiem jelita grubego stwierdzono statystycznie wyższe wartości stężenia interleukiny-2 w porównaniu z innymi grupami • Podwyższony poziom IL-2 w surowicy krwi może być markerem zwiększonego ryzyka raka jelita grubego u osób z cukrzycą typu 2. • Wyniki pracy sugerują, że pacjenci z cukrzycą typu 2 oraz podwyższonym poziomem IL-2 w surowicy krwi powinni być objęci szczególnym nadzorem onkologicznym

Selective Biological Effects of Selenium-Enriched Polysaccharide (Se-Le-30) Isolated from <i>Lentinula edodes</i> Mycelium on Human Immune Cells

A common edible mushroom Lentinula edodes, is an important source of numerous biologically active substances, including polysaccharides, with immunomodulatory and antitumor properties. In the present work, the biological activity of the crude, homogenous (Se)-enriched fraction (named Se-Le-30), which has been isolated from L. edodes mycelium by a modified Chihara method towards human peripheral blood mononuclear cells (PBMCs) and peripheral granulocytes, was investigated. The Se-Le-30 fraction, an analog of lentinan, significantly inhibited the proliferation of human PBMCs stimulated with anti-CD3 antibodies or allostimulated, and down-regulated the production of tumor necrosis factor (TNF)-α by CD3+ T cells. Moreover, it was found that Se-Le-30 significantly reduced the cytotoxic activity of human natural killer (NK) cells. The results suggested the selective immunosuppressive activity of this fraction, which is non-typical for mushroom derived polysaccharides

Interaction between Macrophages and Human Mesenchymal Stromal Cells Derived from Bone Marrow and Wharton’s Jelly—A Comparative Study

Despite intensive clinical research on the use of mesenchymal stromal cells (MSCs), further basic research in this field is still required. Herein, we compared human bone marrow MSCs (BM-MSCs, n = 6) and Wharton’s jelly MSCs (WJ-MSCs, n = 6) in their ability to interact with human primary macrophages. Evaluation of secretory potential revealed that under pro-inflammatory stimulation, WJ-MSCs secreted significantly more IL-6 than BM-MSCs (2-fold). This difference did not translate into the effect of MSCs on macrophages: both types of MSCs significantly directed M1-like macrophages toward the M2 phenotype (based on CD206 expression) to a similar extent. This observation was consistent both in flow cytometry analysis and immunocytochemical assessment. The effect of MSCs on macrophages was sustained when IL-6 signaling was blocked with Tocilizumab. Macrophages, regardless of polarization status, enhanced chemotaxis of both BM-MSCs and WJ-MSCs (p < 0.01; trans-well assay), with WJ-MSCs being significantly more responsive to M1-derived chemotactic signals than BM-MSCs. Furthermore, WJ-MSCs increased their motility (scratch assay) when exposed to macrophage-conditioned medium while BM-MSCs did not. These results indicate that although both BM-MSCs and WJ-MSCs have the ability to reciprocally interact with macrophages, the source of MSCs could slightly but significantly modify the response under clinical settings