hl60 cells  (ATCC)

Journal: Journal of Virology

Article Title: Glycan-reactive antibodies isolated from human HIV-1 vaccine trial participants show broad pathogen cross-reactivity

doi: 10.1128/jvi.01256-25

Figure Lengend Snippet: HVTN 124 mAb ADCP activity. HVTN 124 ADCP activity using native isotypes. ADCP activity was measured against HIV-1 BG505 T332N SOSIP gp140, HIV-1 CNE55 SOSIP gp140, influenza H1N1 A/New Caledonia/30/1999 HA, and SARS-CoV-2 D614G Spike. Antigens were each biotinylated and conjugated to fluorescent neutravidin beads, then mixed with each mAb. Antibodies are listed along the X-axis, while the Y-axis lists the ADCP score as AUC. The control mAbs used include the following: VRC01, PGT128 = HIV-1; Fe53, CR9114 = influenza; CR3022 = SARS-CoV-2. Palivizumab was included as a negative control antibody. Antibodies are color-coded by HVTN124 mAbs (blue), positive control mAbs (green), and negative control mAbs (red). For IgG isotypes, THP-1 monocytes were subsequently added, and ADCP was calculated based on the engulfment of the antigen-coated beads by the THP-1 cells as measured by flow cytometry. For IgA isotypes, DMSO-differentiated HL60 cells were subsequently added, and ADCP was calculated based on the engulfment of the antigen-coated beads by the THP-1 cells as measured by flow cytometry. Antigen engulfment is indicated as an ADCP score. All experiments were performed in triplicate. Additional plots for the native IgA and non-native isotypes can be found in .

Article Snippet: These complexes were then incubated with either THP-1 cells (human monocyte cell line, ATCC TIB-201) for IgG antibodies or DMSO-differentiated HL60 cells (neutrophil-like cell line expressing IgA Fc receptors, ATCC CCL-240) for IgA antibodies.

Techniques: Activity Assay, Control, Negative Control, Positive Control, Flow Cytometry

Journal: bioRxiv

Article Title: Adaptation of Anaplasma phagocytophilum to the tick vector is controlled by the transcriptional regulator Tr1

doi: 10.1101/2025.11.12.688119

Figure Lengend Snippet: tr1 transcription is specific to tick cell infection and necessary for A. phagocytophilum survival in tick cells. ( a ) tr1 transcription normalized to housekeeping gene rpoB during A. phagocytophilum culture in ISE6 tick cells and human HL60 cells. Bar indicate mean of four replicate infections each measured in two technical replicates show as points. ( b and c ) Growth of A. phagocytophilum tr1 ::Himar1 or control strain in cell culture infections of ( b ) human HL60 cells and ( c ) tick ISE6 cells. A. phagocytophilum burden measured by bacterial gDNA relative to eukaryotic host gDNA via qPCR. Data displayed as mean with ±SD of three biological replicates with two technical replicates each. Data are representative of three experimental replicates. * P < 0.05 (Mann-Whitney t -test).

Article Snippet: HL60 human promyelocytic cells (ATCC; CCL-240) were maintained in Roswell Park Memorial Institute (RPMI) 1640 medium with 10% FBS and 1× Glutamax.

Techniques: Infection, Control, Cell Culture, MANN-WHITNEY

Journal: bioRxiv

Article Title: Adaptation of Anaplasma phagocytophilum to the tick vector is controlled by the transcriptional regulator Tr1

doi: 10.1101/2025.11.12.688119

Figure Lengend Snippet: Tr1 binds promotors of neighboring omp1X and omp1N. ( a ) Diagram of tr1 gene with downstream neighboring outer membrane protein genes omp1X , omp1N , and the msp2/p44 expression site. ( b through e ) EMSA shifts with increasing rTr1 (0, 0.0625, 0.125, 0.25, 0.5, and 1μM) with DNA probes of ( b ) tr1 , ( c ) omp1X , ( d ) omp1N , and ( e ) msp2/p44 promotor sequences. ( f and g ) Transcription of omp1X and omp1R from control or tr1 ::Himar1 A. phagocytophilum mutant strains at 24 hours post infection in ( f ) human HL60 or ( g ) tick ISE6 cells. Transcripts measured by qRT-PCR and normalized to rpoB via ΔΔCt. Data displayed as mean with ±SD of four replicate infections measured with two technical replicates each. * P < 0.05 (Mann-Whitney t -test). ( h ) msp2/p44 transcripts totals across all gene variants sequenced by RNAseq from tr1 ::Himar1 or control A. phagocytophilum infected ISE6 tick cells. Bars are mean ±SD of four independent infections, each shown as points. n.s. > 0.05 (Mann-Whitney t -test).

Article Snippet: HL60 human promyelocytic cells (ATCC; CCL-240) were maintained in Roswell Park Memorial Institute (RPMI) 1640 medium with 10% FBS and 1× Glutamax.

Techniques: Membrane, Expressing, Control, Mutagenesis, Infection, Quantitative RT-PCR, MANN-WHITNEY

Journal: bioRxiv

Article Title: Adaptation of Anaplasma phagocytophilum to the tick vector is controlled by the transcriptional regulator Tr1

doi: 10.1101/2025.11.12.688119

Figure Lengend Snippet: Tr1 impacted genes are necessary for survival during tick cell infection. Survival of indicated Himar1 transposon mutant A. phagocytophilum strain relative to intergenic control::Himar1 strain in ( a ) human HL60 (48 hpi) and tick ISE6 cells (8 dpi). Bacterial burden measured by qRT-PCR of A. phagocytophilum 16s vs Ixodes actin transcripts. Data displayed as mean ±SD of four replicate infections measured in two technical replicates each. Graph representative of three replicate experiments. * < 0.005 (Mann-Whitney t -test).

Article Snippet: HL60 human promyelocytic cells (ATCC; CCL-240) were maintained in Roswell Park Memorial Institute (RPMI) 1640 medium with 10% FBS and 1× Glutamax.

Techniques: Infection, Mutagenesis, Control, Quantitative RT-PCR, MANN-WHITNEY

Journal: bioRxiv

Article Title: Adaptation of Anaplasma phagocytophilum to the tick vector is controlled by the transcriptional regulator Tr1

doi: 10.1101/2025.11.12.688119

Figure Lengend Snippet: Tr1 is required for tick-specific remodeling of the T4SS pilus. ( a ) Diagram of the virB2 paralog genes, the location of the virB2_5 ::Himar1 insertion mutation (red), and potential promotor regions subject to EMSA (green). ( b ) Transcription of the virB2 genes during wild-type A. phagocytophilum infection of mammalian HL60 and tick ISE6 cells. Data shown as mean ±SD of four biological replicates. ( c,d,e ) EMSA shifts with increasing rTr1 (0, 0.0625, 0.125, 0.25, 0.5, and 1μM) with putative promotor sequences upstream of ( c ) virB2_1 , ( d ) virB2_6 , and ( e ) virB2_8 . ( f and g ) Growth of A. phagocytophilum virB2_5 ::Himar1 or control strain in cell culture infections of ( f ) human HL60 cells and ( g ) tick ISE6 cells. A. phagocytophilum burden measured by bacterial gDNA relative to eukaryotic host gDNA via qPCR. Data displayed as mean with ±SD of three biological replicates with two technical replicates each. Data are representative of three experimental replicates. * P < 0.05 (Mann-Whitney t -test).

Article Snippet: HL60 human promyelocytic cells (ATCC; CCL-240) were maintained in Roswell Park Memorial Institute (RPMI) 1640 medium with 10% FBS and 1× Glutamax.

Techniques: Mutagenesis, Infection, Control, Cell Culture, MANN-WHITNEY