b lymphocyte cell line raji  (ATCC)

Journal: International Journal of Molecular Sciences

Article Title: Preparation and In Vitro Evaluation of RITUXfab-Decorated Lipoplexes to Improve Delivery of siRNA Targeting C1858T PTPN22 Variant in B Lymphocytes

doi: 10.3390/ijms23010408

Figure Lengend Snippet: Evaluation of FITC-Ritux Fab functionalized liposomes on Raji cells (B-cell lymphoma line). Representative FACS profiles indicating the MFI values of FITC-Ritux Fab among Raji cells. Cells were stained with the anti-CD19-Alexa Fluor 700 ( A ) and FITC-Ritux Fab ( B ) or liposomes functionalized with Ritux Fab /FITC-Ritux Fab (90/10 mol/mol) ( C ).

Article Snippet: The Raji human B lymphocyte cell line (Burkitt’s lymphoma) was obtained from American Type Culture Collection (ATCC CCL-86).

Techniques: Liposomes, Staining

Journal: International Journal of Molecular Sciences

Article Title: Preparation and In Vitro Evaluation of RITUXfab-Decorated Lipoplexes to Improve Delivery of siRNA Targeting C1858T PTPN22 Variant in B Lymphocytes

doi: 10.3390/ijms23010408

Figure Lengend Snippet: Evaluation of targeted lipoplexes on Raji cell line. Representative FACS profiles indicating the percentages of Fab(CD20)-FITC+ among Raji cells after staining with unlabeled and untargeted lipoplex ( A ), FITC-Ritux Fab ( B ) or targeted lipoplex ( C ).

Article Snippet: The Raji human B lymphocyte cell line (Burkitt’s lymphoma) was obtained from American Type Culture Collection (ATCC CCL-86).

Techniques: Staining

Journal: eLife

Article Title: Perinatal hormones favor CC17 group B Streptococcus intestinal translocation through M cells and hypervirulence in neonates

doi: 10.7554/eLife.48772

Figure Lengend Snippet: Human Caco-2 enterocytes were polarized on standard plates ( a ) or Transwell inserts ( b to h ) for 14–21 days. ( b to h ) Human Raji B lymphocytes were added in the lower compartment of the culture chamber in order to obtain a cellular layer composed of enterocytes and M cells and the cell culture medium was supplemented with E2-P4 concentrations equivalent to those found at birth (E2: 10 −8 M, P4: 10 −6 M; E2-P4 C 0 condition) and 7 days later (E2: 10 −9 M, P4: 10 −7 M; E2-P4 C 7 condition) for 6 days before infection. Control cells were maintained in regular cell culture medium without E2 and P4 supplementation. Epithelial cells were infected by their apical side with bacteria at a multiplicity of infection 10 ( a ) and 100 ( c to h ) for 2 hr. ( a ) Bacterial adherence and invasion of a cellular monolayer composed of enterocytes. ( b ) Representative confocal microscopy image of the epithelial layer obtained following Caco-2 and Raji co-culture showing enterocytes and M cells. ( c ) Transcytosis of CC17 GBS across a monolayer strictly composed of enterocytes and a monolayer composed of both enterocytes and M cells. ( d ) Representative confocal microscopy images showing CC17 and CC23 GBS adherence to the enterocytes + M cells epithelial layer. ( e ) Bacterial adherence of CC17 and CC23 GBS to the enterocytes + M cells epithelial layer. ( f ) Quantitative imaging analysis of bacterial association with M cells expressed as the proportion of bacteria associated with GP2 positive cells relative to total adherent bacteria. ( g ) Representative confocal microscopy image showing the presence of several CC17 GBS cocci at the surface and inside an M cell. ( g1 to g3 ) Orthogonal views of ( g ). ( d to g ) Experiments were carried out in E2-P4 C 7 condition. ( h ) Quantitative imaging analysis of CC17 GBS association with M cells in E2-P4 C 0 , E2-P4 C 7 , and control condition. ( a, c, e, f, h ) Results are expressed as mean ± SEM.≥3 experiments in duplicate. ( b, d, g ) Tight junctions are labeled with anti-ZO1 antibody, M cells with anti-GP2 antibody, GBS with anti-GBS antibody, and actin with phalloidin. Multiple-group comparisons were performed by Kruskal-Wallis test ( c and h ) and single-group comparisons by Mann-Whitney test ( e and f ). *p<0.05; **p<0.01, ***p<0.001.

Article Snippet: The human B lymphocytes cell line Raji (ATCC CCL-86, RRID: CVCL_0511 ) was cultured at 37°C in a 5% CO 2 atmosphere in RPMI 1640 medium with GlutaMax (ThermoFisher) with 10% inactivated FCS (Eurobio).

Techniques: Cell Culture, Infection, Control, Bacteria, Confocal Microscopy, Co-Culture Assay, Imaging, Labeling, MANN-WHITNEY

Journal: eLife

Article Title: Perinatal hormones favor CC17 group B Streptococcus intestinal translocation through M cells and hypervirulence in neonates

doi: 10.7554/eLife.48772

Figure Lengend Snippet: Human Caco-2 enterocytes were polarized on Transwell inserts for 14–21 days. Human Raji B lymphocytes were added in the lower compartment of the culture chamber for the six following days in order to obtain a cellular layer composed of enterocytes and M cells. The cell culture medium was supplemented with E2-P4 concentrations equivalent to those found at birth (E2: 10 −8 M, P4: 10 −6 M; E2-P4 C 0 condition) and 7 days later (E2: 10 −9 M, P4: 10 −7 M; E2-P4 C 7 condition) for 6 days before infection. Epithelial cells were infected by their apical side with bacteria at a multiplicity of infection 100 for 2 hr. Representative confocal microscopy images (out of 10 per experiment) of CC17 and CC23 GBS association with the enterocytes + M cells epithelial layer in E2-P4 C 7 condition. GBS were labeled with anti-GBS antibody, M cells with anti-GP2 antibody, actin with phalloidin, and nuclei with DAPI.

Article Snippet: The human B lymphocytes cell line Raji (ATCC CCL-86, RRID: CVCL_0511 ) was cultured at 37°C in a 5% CO 2 atmosphere in RPMI 1640 medium with GlutaMax (ThermoFisher) with 10% inactivated FCS (Eurobio).

Techniques: Cell Culture, Infection, Bacteria, Confocal Microscopy, Labeling

Journal: eLife

Article Title: Perinatal hormones favor CC17 group B Streptococcus intestinal translocation through M cells and hypervirulence in neonates

doi: 10.7554/eLife.48772

Figure Lengend Snippet: ( a–d ) The epithelial layer composed of enterocytes and M cells was prepared as described in . Following 6 days of Caco-2 and Raji co-culture, lymphocytes and epithelial cells were recovered and lysed for gene and protein expression analysis. ( a ) Raji B lymphocytes mRNA levels of RANKL. ( b ) Caco-2 mRNA levels of Spi-B-dependent M cells differentiation genes. ( a–b ) Results are normalized to ACTIN and expressed as mean fold change relative to control condition ± SEM. ( c–d ) Western blot analysis of GP2. ( c ) Representative sample blot and ( d ) quantification of GP2 protein levels. Values are presented as the ratio between GP2 and Actin and expressed as mean fold change relative to control condition ± SEM.≥3 experiments in duplicate. ( e ) SPF 3-week-old mice were administered E2-P4 cocktails as described in . At the end of the hormonal treatment, Peyer’s patches were recovered for mRNA quantification of M cells differentiation markers. Results are normalized to Actin and expressed as mean fold change relative to control condition ± SEM (n = 10 mice per group). *p<0.05, **p<0.01, ***p<0.001, ns: not significant (Kruskal-Wallis test).

Article Snippet: The human B lymphocytes cell line Raji (ATCC CCL-86, RRID: CVCL_0511 ) was cultured at 37°C in a 5% CO 2 atmosphere in RPMI 1640 medium with GlutaMax (ThermoFisher) with 10% inactivated FCS (Eurobio).

Techniques: Co-Culture Assay, Expressing, Control, Western Blot

Journal: eLife

Article Title: Perinatal hormones favor CC17 group B Streptococcus intestinal translocation through M cells and hypervirulence in neonates

doi: 10.7554/eLife.48772

Figure Lengend Snippet:

Article Snippet: The human B lymphocytes cell line Raji (ATCC CCL-86, RRID: CVCL_0511 ) was cultured at 37°C in a 5% CO 2 atmosphere in RPMI 1640 medium with GlutaMax (ThermoFisher) with 10% inactivated FCS (Eurobio).

Techniques: Isolation, Construct, Plasmid Preparation, Software