Journal: Scientific Reports

Article Title: The contribution of the membrane-bound complement regulatory proteins CD46 and CD55 in phases of acute lymphocytic leukemia and acute myelogenous leukemia

doi: 10.1038/s41598-025-33359-y

Figure Lengend Snippet: CD46 and CD55 mRNA expression in acute leukemia patients. CD46 and CD55 mRNA expression levels were measured using qRT-PCR in both AML and ALL patients, normalized to the housekeeping gene Beta-actin and results were compared to CD46 and CD55 mRNA expression levels in healthy controls by calculating the RQ value for each one of them. mRNA expression of mCRPs was significantly reduced in AML and ALL patients in comparison with healthy controls. Data are expressed as means ± SD of three independent experiments for each sample done in duplicates and statistical one-way analysis of variance (ANOVA) was used to estimate the significance of difference between the different groups. P < 0.001(***).

Article Snippet: Gene expression analysis for CD46 and CD55 was done using Taqman gene expression assays Hs00611257_m1 and Hs00892618_m1 (Thermofisher scientific, USA), respectively, with Beta Actin (Hs01060665 g1) being the housekeeping or reference gene.

Techniques: Expressing, Quantitative RT-PCR, Comparison

Journal: Scientific Reports

Article Title: The contribution of the membrane-bound complement regulatory proteins CD46 and CD55 in phases of acute lymphocytic leukemia and acute myelogenous leukemia

doi: 10.1038/s41598-025-33359-y

Figure Lengend Snippet: CD55 mRNA expression level based on gender differences. The expression level of CD55 in males and females showed a slight reduction in female AML patients compared to males, while the opposite was observed in ALL patients. Data are expressed as means ± SD of three independent experiments for each sample done in duplicates and statistical one-way analysis of variance (ANOVA) was used to estimate the significance of difference between the different groups.

Article Snippet: Gene expression analysis for CD46 and CD55 was done using Taqman gene expression assays Hs00611257_m1 and Hs00892618_m1 (Thermofisher scientific, USA), respectively, with Beta Actin (Hs01060665 g1) being the housekeeping or reference gene.

Techniques: Expressing

Journal: Scientific Reports

Article Title: The contribution of the membrane-bound complement regulatory proteins CD46 and CD55 in phases of acute lymphocytic leukemia and acute myelogenous leukemia

doi: 10.1038/s41598-025-33359-y

Figure Lengend Snippet: Flow cytometric analysis of the relative expression level of CD46 and CD55 proteins in AML and ALL patients. Flow cytometric analysis performed to measure the expression level of CD46 and CD55 in AML and ALL patients showed a significant reduction in CD46 expression in both types of acute leukemia patients. However, for CD55 a significant reduction was observed in AML patients only, while there was no significant difference observed in case of ALL patients. The expression level was calculated with reference to healthy controls. Data are represented as mean values ± SD for each sample done in duplicates, p < 0.05 (*).

Article Snippet: Gene expression analysis for CD46 and CD55 was done using Taqman gene expression assays Hs00611257_m1 and Hs00892618_m1 (Thermofisher scientific, USA), respectively, with Beta Actin (Hs01060665 g1) being the housekeeping or reference gene.

Techniques: Expressing

Journal: Scientific Reports

Article Title: The contribution of the membrane-bound complement regulatory proteins CD46 and CD55 in phases of acute lymphocytic leukemia and acute myelogenous leukemia

doi: 10.1038/s41598-025-33359-y

Figure Lengend Snippet: Flow cytometric analysis charts of CD46 and CD55 in healthy controls and acute leukemia patients’ peripheral blood samples. Flow cytometric analysis of the protein expression level of both CD46 and CD55 in the peripheral blood samples of acute leukemia patients showed a significant reduction in both proteins compared to healthy controls. Results shown are from a representative experiment. ( A ) CD46 expression in a healthy control. ( B ) CD46 expression in an AML patient. ( C ) CD46 expression in an ALL patient. ( D ) CD55 expression in a healthy control. ( E ) CD55 expression in an AML patient. ( F ) CD55 expression in an ALL patient.

Article Snippet: Gene expression analysis for CD46 and CD55 was done using Taqman gene expression assays Hs00611257_m1 and Hs00892618_m1 (Thermofisher scientific, USA), respectively, with Beta Actin (Hs01060665 g1) being the housekeeping or reference gene.

Techniques: Expressing, Control

Journal: Scientific Reports

Article Title: The contribution of the membrane-bound complement regulatory proteins CD46 and CD55 in phases of acute lymphocytic leukemia and acute myelogenous leukemia

doi: 10.1038/s41598-025-33359-y

Figure Lengend Snippet: ShRNA-mediated knockdown of CD46 and CD55 mCRPs expression on HSB-2 cells compared to the mock transfected controls. Knockdown of CD46 and CD55 protein expression in transfected cells was performed using shRNA silencing plasmids, each consisting of a pool of three different shRNA plasmids and PEI was used as the transfection reagent. Silencing of CD46 and CD55 proteins was done separately in addition to co-silencing of both proteins. Flow cytometric analysis was performed to measure the expression level of both proteins post transfection. Results showed a highly significant reduction in the expression level of both proteins. The percentage of inhibition was calculated relative to nonsilencing shRNA controls (= 100%) p < 0.001(***). ( A ) CD46 and CD55 proteins expression level post transfection compared to expression in mock transfected cells. ( B ) Flow cytometric analysis chart of a representative experiment showing protein expression of CD46 in HSB-2 transfected cells. ( C ) Flow cytometric analysis chart of a representative experiment showing protein expression of CD55 in HSB-2 transfected cells. ( D ) Flow cytometric analysis chart of a representative experiment showing protein expression of CD46 and CD55 in mock transfected cells.

Article Snippet: Gene expression analysis for CD46 and CD55 was done using Taqman gene expression assays Hs00611257_m1 and Hs00892618_m1 (Thermofisher scientific, USA), respectively, with Beta Actin (Hs01060665 g1) being the housekeeping or reference gene.

Techniques: shRNA, Knockdown, Expressing, Transfection, Inhibition

Journal: Scientific Reports

Article Title: The contribution of the membrane-bound complement regulatory proteins CD46 and CD55 in phases of acute lymphocytic leukemia and acute myelogenous leukemia

doi: 10.1038/s41598-025-33359-y

Figure Lengend Snippet: ShRNA-mediated combined knockdown of CD46 and CD55 mCRPs expression on HSB-2 cells compared to the mock transfected controls. ( A ) Co-silencing of CD46 and CD55 expression in transfected cells led to a highly significant reduction in the expression of both proteins( p < 0.001). A significant difference was observed between CD46 and CD55 knockdown( p < 0.05).The percentage of inhibition was calculated relative to nonsilencing shRNA controls (= 100%). p < 0.001(***), p < 0.05(*). ( B ) Flow cytometry analysis chart for CD46 and CD55 expression on HSB-2 cells transfected with combined CD46 and CD55 shRNA silencing plasmids. Dots in the third quadrant show the double negative expression for both CD46 and CD55 after knockdown of both CD46 and CD55 genes using combined CD46 and CD55 silencing shRNA plasmids.

Article Snippet: Gene expression analysis for CD46 and CD55 was done using Taqman gene expression assays Hs00611257_m1 and Hs00892618_m1 (Thermofisher scientific, USA), respectively, with Beta Actin (Hs01060665 g1) being the housekeeping or reference gene.

Techniques: shRNA, Knockdown, Expressing, Transfection, Inhibition, Flow Cytometry

Journal: Scientific Reports

Article Title: The contribution of the membrane-bound complement regulatory proteins CD46 and CD55 in phases of acute lymphocytic leukemia and acute myelogenous leukemia

doi: 10.1038/s41598-025-33359-y

Figure Lengend Snippet: Acute lymphocytic leukemia cells viability after silencing of mCRP. Viability of HSB-2 cells was analyzed after silencing of CD46 and CD55 protein expression separately as well as combined silencing of both proteins using shRNA plasmids. Cell viability was assessed using MTT reagent in the presence of NHS as a source of complement to mimic exposure to physiological complement. Cells had a reduced viability in the absence of CD46 ( p < 0.001) and CD55 ( p < 0.01) as well as in CD46 and CD55 co-silenced cells ( p < 0.01). These results demonstrate that CD46 and CD55 contribute to the protection of leukemic cells against complement attack, and that their complete depletion sensitizes cells to CDC, which directly links this assay to their contribution in immune escape. P < 0.01 (**), p < 0.001 (***).

Article Snippet: Gene expression analysis for CD46 and CD55 was done using Taqman gene expression assays Hs00611257_m1 and Hs00892618_m1 (Thermofisher scientific, USA), respectively, with Beta Actin (Hs01060665 g1) being the housekeeping or reference gene.

Techniques: Expressing, shRNA