Alterations in TP53, cyclin D2, c-Myc, p21WAF1/CIP1 and p27KIP1 expression associated with progression in B-CLL.

B-cell chronic lymphocytic leukaemia (B-CLL) originates from B lymphocytes that may differ in the activation level, maturation state or cellular subgroups in peripheral blood. Tumour progression in CLL B cells seems to result in gradual accumulation of the clone of resting B lymphocytes in the early phases (G0/G1) of the cell cycle. The G1 phase is impaired in B-CLL. We investigated the gene expression of five key cell cycle regulators: TP 53, c-Myc, cyclin D2, p21WAF1/CIP1 and p27KIP1, which primarily regulate the G1 phase of the cell cycle, or S-phase entry and ultimately control the proliferation and cell growth as well as their role in B-CLL progression. The study was conducted in peripheral blood CLL lymphocytes of 40 previously untreated patients. Statistical analysis of correlations of TP53, cyclin D2, c-Myc, p21WAF1/CIP1 and p27KIP1 expressions in B-CLL patients with different Rai stages demonstrated that the progression of disease was accompanied by increases in p53, cyclin D2 and c-Myc mRNA expression. The expression of p27KIP1 was nearly statistically significant whereas that of p21 WAF1/CIP1 showed no such correlation. Moreover, high expression levels of TP53 and c-Myc genes were found to be closely associated with more aggressive forms of the disease requiring earlier therapy

Impaired endotoxin tolerance on example of IL6 in B-CLL

IL-6 uznaje się za kluczową cytokinę zaangażowaną w mechanizmy obronne organizmu, w procesach krwiotworzenia i reakcjach zapalnych, o wielokierunkowym działaniu auto- i parakrynnym. Jej produkcja jest zazwyczaj przemijająca i ściśle regulowana. Stosując metodę RT-PCR w czasie rzeczywistym (real time PCR) zbadano ekspresję IL-6 na poziomie mRNA, w prawidłowych i białaczkowych limfocytach B spoczynkowych i stymulowanych LPS przez 30' i 24 h. Poziom ekspresji IL-6 wyrażony wartością RQ w puli limfocytów PBL-B, w porównaniu z subpopulacją limfocytów B zdrowych dawców, był istotnie statystycznie niższy, wartość p=0,00348. Po 24 h w prawidłowych limfocytach B poziom RQ mRNA IL-6 istotnie statystycznie obniżał się (p=0,012970), natomiast w subpopulacji PBL-B istotnie statystycznie nadal wzrastał (p=0,000001). Uzyskane dane wydają się wskazywać na zaburzanie tolerancji endotoksycznej w PBL-BIL-6 is considered a key cytokine involved in immune organism response, hematopoietic processes and inflammatory reactions, based on multidirectional auto and paracrine auction Its production is usually transient and tightly regulated.By the means of Real Time PCR we studied IL-6 expression on mRNA level (RQ) in B-CLL lymphocytes and in CD19+ subpopulation of normal B lymphocytes before and after 30 min and 24h LPS stimulation. The IL-6 expression level (RQ) in B-CLL cells was significantly lower (p=0.00348) compared with the subpopulation of CD19+ normal lymphocytes. 30 min LPS stimulation resulted in increase of IL6 mRNA expression, both in normal and B-CLL lymphocytes. IL6 mRNA expression level decreased after 24h LPS stimulation (p=0.012970) in normal lymphocytes, whereas B-CLL cells showed constant increase (p=0.000001). These data seem to indicate an aberrant endotoxin tolerance in B-CLL

NOD1 and NOD2 receptors: integral members of the innate and adaptive immunity system

NOD-like proteins (NLR) are a specialized group of intracellular receptors, which constitute an essential component of the host innate immune system. They were discovered more than a decade ago, but research on this particular class of microbial detectors is still ongoing to allow for a better understanding of the mechanisms, recognition of microorganisms, transmission of signals, and carrying out the activation of inflammatory signaling pathways. In this review, we discuss the construction of NOD1 and NOD2 receptors, their functions, and significance in the pathogenesis of inflammatory diseases in humans

Different expression of CD180, CD284 and CD14 receptors on the CD19+ subpopulation of normal and B-CLL lymphocytes.

Numerous experimental data indicate that B-CLL development and progression are influenced by antigenic pressure. It can not be excluded that these antigens may originate from bacteria and viruses. Toll like receptors (TLRs) interact with pathogen associated molecular patterns as part of innate immunity. TLRs are currently used to target different subclasses of B-cell leukemia, and TLR agonists are being evaluated in clinical trials. It is little known regarding the repertoire and function of TLR in B-CLL. The aim of the study was to assess the CD180, CD284 and mCD14 levels in CD19+ subpopulation of B-CLL peripheral blood lymphocytes and compare them with respective levels in the normal B-cells of adult volunteers, before and after LPS stimulation. We investigated the percentage of the CD19+CD180+, CD19+CD284+, CD19+CD14+ cells and the mean fluorescence intensity (MFI) of CD180, CD284 and CD14 antigens among CD19+ B-CLL as well as in the normal B cells for comparison. MFI analysis revealed that CD180, CD284 and CD14 expression was higher on normal B cells then on CD19+ B-CLL (MFI CD180: 99.16 vs. 25.3, MFI CD284: 7.37 vs. 5.79 and MFI CD14 25.07 vs. 8.32). After 24-hour LPS activation of B-cells, CD180 MFI appeared to decrease, in both healthy and B-CLL patients. CD284 MFI in healthy controls decreased after LPS stimulation while slight increase of MFI was observed in leukemic cells. CD14 MFI in leukemic cells was moderately higher after LPS in comparison to CD14 MFI without LPS stimulation, whereas CD14 MFI in normal CD19+ cells after LPS stimulation decreased over three times. Variations observed in expression of both normal and leukemic receptors may be due to their different sensitivity to antigenic stimulation

Ekspresja genu podoplaniny w złośliwych nowotworach jajnika

Introduction: Ovarian cancer is mostly diagnosed in postmenopausal women. The hormonal microenvironmentof ovarian tumour development in pre- and postmenopausal women is different. The environment mayinfluence the molecular basis of ovarian cancer. Podoplanin is a proven marker of lymphangiogenesis and isengaged in tumour progression and metastasis. Objectives: The aim of our study was to assess podoplanin expression at the mRNA level in ovarian serousand mucinous adenocarcinoma. Material and methods: mRNA expression of podoplanin was assessed in 42 ovarian cancer patients bymeans of real-time PCR. Results: Podoplanin was expressed at the mRNA level in all ovarian cancer patients. It was significantlylower (p < 0.001) in cancer tissues (RQ = 4.46) than in healthy control (RQ = 13.03). No significant differenceswere found in podoplanin expression either between pre- and postmenopausal women or in relation to FIGOand grading. Conclusion: Decreased podoplanin expression is characteristic for serous and mucinous ovarianadenocarcinoma

Different expression of CD180, CD284 and CD14 receptors on the CD19+ subpopulation of normal and B-CLL lymphocytes.

Numerous experimental data indicate that B-CLL development and progression are influenced by antigenic pressure. It can not be excluded that these antigens may originate from bacteria and viruses. Toll like receptors (TLRs) interact with pathogen associated molecular patterns as part of innate immunity. TLRs are currently used to target different subclasses of B-cell leukemia, and TLR agonists are being evaluated in clinical trials. It is little known regarding the repertoire and function of TLR in B-CLL. The aim of the study was to assess the CD180, CD284 and mCD14 levels in CD19+ subpopulation of B-CLL peripheral blood lymphocytes and compare them with respective levels in the normal B-cells of adult volunteers, before and after LPS stimulation. We investigated the percentage of the CD19+CD180+, CD19+CD284+, CD19+CD14+ cells and the mean fluorescence intensity (MFI) of CD180, CD284 and CD14 antigens among CD19+ B-CLL as well as in the normal B cells for comparison. MFI analysis revealed that CD180, CD284 and CD14 expression was higher on normal B cells then on CD19+ B-CLL (MFI CD180: 99.16 vs. 25.3, MFI CD284: 7.37 vs. 5.79 and MFI CD14 25.07 vs. 8.32). After 24-hour LPS activation of B-cells, CD180 MFI appeared to decrease, in both healthy and B-CLL patients. CD284 MFI in healthy controls decreased after LPS stimulation while slight increase of MFI was observed in leukemic cells. CD14 MFI in leukemic cells was moderately higher after LPS in comparison to CD14 MFI without LPS stimulation, whereas CD14 MFI in normal CD19+ cells after LPS stimulation decreased over three times. Variations observed in expression of both normal and leukemic receptors may be due to their different sensitivity to antigenic stimulation