Journal: International Journal of Molecular Sciences

Article Title: gp120 Envelope Glycoproteins of HIV-1 Group M Subtype A and Subtype B Differentially Affect Gene Expression in Human Vascular Endothelial Cells

doi: 10.3390/ijms24043536

Figure Lengend Snippet: Characteristics of recombinant proteins used in this study.

Article Snippet: Cells were treated in triplicate with either the recombinant gp120 of HIV-1 Group M Subunit A (catalog number 40403-V08H, Sino Biological, Inc., Beijing, China), gp120 of HIV-2 Group M Subunit B (catalog number 40404-V08H, Sino Biological), S1 of SARS CoV-2 spike protein (catalog number 40591-V08H, Sino Biological), or S1 of the omicron variant SARS CoV-2 spike protein (catalog number 40591-V08H41, Sino Biological).

Techniques: Recombinant, Sequencing, Expressing, Variant Assay

Journal: International Journal of Molecular Sciences

Article Title: gp120 Envelope Glycoproteins of HIV-1 Group M Subtype A and Subtype B Differentially Affect Gene Expression in Human Vascular Endothelial Cells

doi: 10.3390/ijms24043536

Figure Lengend Snippet: gp120 proteins of HIV-1 Group M Subtype A and Subtype B differentially affect prostasin, MMP-2, and ErbB3 expression. Human pulmonary artery endothelial cells were treated with gp120 of HIV-1 Subtype A (gp120A) or Subtype B (gp120B) at 1 nM for 20 h in triplicate. Expression patterns of various proteins were monitored using R&D Human XL Oncology Array. ( A ) Prostasin was found to be higher in gp120B-treated cells, while neighboring E-selectin spots were unchanged. ( B ) MMP-2 was found to be higher in gp120B-treated cells, while neighboring progranulin spots were unchanged. ( C ) ErbB3 was found to be higher in gp120B-treated cells, while neighboring Dkk-1 spots were unchanged. Representative images are shown at the top. Densitometry values from two spots from each array were averaged, and statistical analysis was performed using results from three separate treatments/arrays. Bar graphs represent means ± SEM (N = 3 for all groups). * p < 0.05. ** p < 0.01. *** p < 0.001. The y -axis indicates the mean pixel density of protein spots.

Article Snippet: Cells were treated in triplicate with either the recombinant gp120 of HIV-1 Group M Subunit A (catalog number 40403-V08H, Sino Biological, Inc., Beijing, China), gp120 of HIV-2 Group M Subunit B (catalog number 40404-V08H, Sino Biological), S1 of SARS CoV-2 spike protein (catalog number 40591-V08H, Sino Biological), or S1 of the omicron variant SARS CoV-2 spike protein (catalog number 40591-V08H41, Sino Biological).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: gp120 Envelope Glycoproteins of HIV-1 Group M Subtype A and Subtype B Differentially Affect Gene Expression in Human Vascular Endothelial Cells

doi: 10.3390/ijms24043536

Figure Lengend Snippet: Characteristic of the effects of gp120 proteins of HIV-1 Group M Subtype A and Subtype B on the prostasin expression. Human pulmonary artery endothelial cells were treated with SARS-CoV-2 spike proteins S1, Omicron S1, and gp120 of HIV-1 Subtype A (gp120A) or Subtype B (gp120B) at 1 nM for 20 h in triplicate. Expression patterns of various proteins were monitored using R&D Human XL Oncology Array. Bar graphs represent means ± SEM (N = 3 for all groups) of ( A ) prostasin expression, ( B ) E-selectin expression, and ( C ) the ratio of prostasin to E-selectin expression. * p < 0.05. ** p < 0.01. *** p < 0.001. The y -axis indicates the mean pixel density of protein spots.

Article Snippet: Cells were treated in triplicate with either the recombinant gp120 of HIV-1 Group M Subunit A (catalog number 40403-V08H, Sino Biological, Inc., Beijing, China), gp120 of HIV-2 Group M Subunit B (catalog number 40404-V08H, Sino Biological), S1 of SARS CoV-2 spike protein (catalog number 40591-V08H, Sino Biological), or S1 of the omicron variant SARS CoV-2 spike protein (catalog number 40591-V08H41, Sino Biological).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: gp120 Envelope Glycoproteins of HIV-1 Group M Subtype A and Subtype B Differentially Affect Gene Expression in Human Vascular Endothelial Cells

doi: 10.3390/ijms24043536

Figure Lengend Snippet: Characteristic of the effects of gp120 proteins of HIV-1 Group M Subtype A and Subtype B on the MMP-2 expression. Human pulmonary artery endothelial cells were treated with SARS-CoV-2 spike proteins S1, Omicron S1, and gp120 of HIV-1 Subtype A (gp120A) or Subtype B (gp120B) at 1 nM for 20 h in triplicate. Expression patterns of various proteins were monitored using R&D Human XL Oncology Array. Bar graphs represent means ± SEM (N = 3 for all groups) of ( A ) MMP-2 expression, ( B ) progranulin expression, and ( C ) the ratio of MMP-2 to progranulin expression. * p < 0.05. ** p < 0.01. *** p < 0.001. The y -axis indicates the mean pixel density of protein spots.

Article Snippet: Cells were treated in triplicate with either the recombinant gp120 of HIV-1 Group M Subunit A (catalog number 40403-V08H, Sino Biological, Inc., Beijing, China), gp120 of HIV-2 Group M Subunit B (catalog number 40404-V08H, Sino Biological), S1 of SARS CoV-2 spike protein (catalog number 40591-V08H, Sino Biological), or S1 of the omicron variant SARS CoV-2 spike protein (catalog number 40591-V08H41, Sino Biological).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: gp120 Envelope Glycoproteins of HIV-1 Group M Subtype A and Subtype B Differentially Affect Gene Expression in Human Vascular Endothelial Cells

doi: 10.3390/ijms24043536

Figure Lengend Snippet: Characteristic of the effects gp120 proteins of HIV-1 Group M Subtype A and Subtype B on the ErbB3 expression. Human pulmonary artery endothelial cells were treated with SARS-CoV-2 spike proteins S1, Omicron S1, and gp120 of HIV-1 Subtype A (gp120A) or Subtype B (gp120B) at 1 nM for 20 h in triplicate. Expression patterns of various proteins were monitored using R&D Human XL Oncology Array. Bar graphs represent means ± SEM (N = 3 for all groups) of ( A ) ErbB3 expression, ( B ) Dkk-1 expression, and ( C ) the ratio of ErbB3 to Dkk-1 expression. * p < 0.05. ** p < 0.01. *** p < 0.001. The y -axis indicates the mean pixel density of protein spots.

Article Snippet: Cells were treated in triplicate with either the recombinant gp120 of HIV-1 Group M Subunit A (catalog number 40403-V08H, Sino Biological, Inc., Beijing, China), gp120 of HIV-2 Group M Subunit B (catalog number 40404-V08H, Sino Biological), S1 of SARS CoV-2 spike protein (catalog number 40591-V08H, Sino Biological), or S1 of the omicron variant SARS CoV-2 spike protein (catalog number 40591-V08H41, Sino Biological).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: gp120 Envelope Glycoproteins of HIV-1 Group M Subtype A and Subtype B Differentially Affect Gene Expression in Human Vascular Endothelial Cells

doi: 10.3390/ijms24043536

Figure Lengend Snippet: gp120 proteins of HIV-1 Group M Subtype A and Subtype B differentially affect MCP-2, and MCP-3 expression. Human pulmonary artery endothelial cells were treated with SARS-CoV-2 spike proteins S1, Omicron S1, and gp120 of HIV-1 Subtype A (gp120A) or Subtype B (gp120B) at 1 nM for 20 h in triplicate. Expression patterns of various proteins were monitored using R&D Human XL Oncology Array. ( A ) MCP-2 was found to be higher in gp120A-treated cells. ( B ) MMP-3 was found to be higher in gp120A-treated cells. Representative images are shown at the top. Densitometry values from two spots from each array were averaged, and statistical analysis was performed using results from three separate treatments/arrays. Bar graphs represent means ± SEM (N = 3 for all groups). * p < 0.05. ** p < 0.01. The y -axis indicates the mean pixel density of protein spots.

Article Snippet: Cells were treated in triplicate with either the recombinant gp120 of HIV-1 Group M Subunit A (catalog number 40403-V08H, Sino Biological, Inc., Beijing, China), gp120 of HIV-2 Group M Subunit B (catalog number 40404-V08H, Sino Biological), S1 of SARS CoV-2 spike protein (catalog number 40591-V08H, Sino Biological), or S1 of the omicron variant SARS CoV-2 spike protein (catalog number 40591-V08H41, Sino Biological).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: gp120 Envelope Glycoproteins of HIV-1 Group M Subtype A and Subtype B Differentially Affect Gene Expression in Human Vascular Endothelial Cells

doi: 10.3390/ijms24043536

Figure Lengend Snippet: gp120 proteins of HIV-1 Group M Subtype A and Subtype B differentially affect TARC expression. Human pulmonary artery endothelial cells were treated with gp120 of HIV-1 Subtype A (gp120A) or Subtype B (gp120B) at 1 nM for 20 h. Expression patterns of various proteins were monitored using R&D Human XL Cytokine Array. TARC expression was found to be higher in gp120A-treated cells, while neighboring angiopoietin-1 spots were unchanged. Representative images are shown at the top. Densitometry values from two spots from each array were averaged, and statistical analysis was performed using results from three separate treatments/arrays. Bar graphs represent means ± SEM (N = 3 for gp120A and gp120B groups; N = 2 for untreated control). * p < 0.05. *** p < 0.001. The y -axis indicates the mean pixel density of protein spots.

Article Snippet: Cells were treated in triplicate with either the recombinant gp120 of HIV-1 Group M Subunit A (catalog number 40403-V08H, Sino Biological, Inc., Beijing, China), gp120 of HIV-2 Group M Subunit B (catalog number 40404-V08H, Sino Biological), S1 of SARS CoV-2 spike protein (catalog number 40591-V08H, Sino Biological), or S1 of the omicron variant SARS CoV-2 spike protein (catalog number 40591-V08H41, Sino Biological).

Techniques: Expressing

Journal: bioRxiv

Article Title: Multiple mechanisms of self-association of chemokine receptors CXCR4 and CCR5 demonstrated by deep mutagenesis

doi: 10.1101/2023.03.25.534231

Figure Lengend Snippet: Formation of syncytia was measured by flow cytometry after co-culturing X4-KO Expi293F cells expressing either ( A ) MN gp160 or ( B ) BaL gp160 with cells expressing CD4 and the indicated CXCR4 or CCR5 mutants, respectively. Data are mean ± SD, n = 6 independent experiments. *, p < 0.05; **, p < 0.01, Student’s t test.

Article Snippet: Plasmids for HIV-1 MN gp160 and BaL gp160 (# 100919) are previously described ( ). pCMV3-CD4 was from Sino Biological (# HG10400-UT).

Techniques: Flow Cytometry, Expressing

Journal:

Article Title: Prime-Boost Immunization Schedules Based on Influenza Virus and Vaccinia Virus Vectors Potentiate Cellular Immune Responses against Human Immunodeficiency Virus Env Protein Systemically and in the Genitorectal Draining Lymph Nodes

doi: 10.1128/JVI.77.12.7048-7057.2003

Figure Lengend Snippet: Evaluation of specific cellular immune responses against HIV-1 Env antigen. Quantification of Env peptide specific IFN-γ-secreting CD8+ T cells. Four BALB/c mice per group were first intranasally (i.n.) immunized with 104 PFU of influenza virus Env, and 14 days later the animals were intranasally boosted with 104 PFU of influenza virus Env or intraperitoneally (i.p.) boosted with 107 PFU of VV WR Env or MVA Env. Cell suspensions of the spleens (A) or genitorectal lymph nodes (B) obtained 14 days after the immunization were evaluated for Env-specific IFN-γ-secreting cells by ELISPOT assay. The number of antigen-specific IFN-γ-secreting cells with standard deviation from triplicate cultures is shown. The pattern of cytokine secretion after gp160 restimulation of cell suspensions of spleens (C) or genitorectal lymph nodes (D) was determined. After 72 h of culture, cell culture supernatants were harvested and evaluated for IFN-γ and IL-4 by ELISA. Bars represent the median ± standard deviation of triplicate samples.

Article Snippet: Lymphocytes were cultured in triplicate (10 6 cells/well) in 96-well microtiter flat-bottomed plates and stimulated with purified gp160 protein (Intracel Corporation, Cambridge, Mass.) (1 μg/ml) or concanavalin A (1 μg/ml) (Sigma Chemical).

Techniques: Enzyme-linked Immunospot, Standard Deviation, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal:

Article Title: Prime-Boost Immunization Schedules Based on Influenza Virus and Vaccinia Virus Vectors Potentiate Cellular Immune Responses against Human Immunodeficiency Virus Env Protein Systemically and in the Genitorectal Draining Lymph Nodes

doi: 10.1128/JVI.77.12.7048-7057.2003

Figure Lengend Snippet: Induction of anti-V3 Env antibodies and IgG2a/IgG1 ratios after immunization with influenza virus and VV vectors. Sera from mice of the experiment described in Fig. ​Fig.11 were evaluated for specific anti-gp160 antibodies 14 days after booster. Reactivity of individual serum samples against V3 peptide was assayed by ELISA. (A) Absorbances for IgG1 and IgG2a subclasses in 1:50 serum dilutions from mice of the different groups (absorbance from sera of nonimmunized mice was subtracted). (B) IgG2a/IgG1 ratios in the different groups.

Article Snippet: Lymphocytes were cultured in triplicate (10 6 cells/well) in 96-well microtiter flat-bottomed plates and stimulated with purified gp160 protein (Intracel Corporation, Cambridge, Mass.) (1 μg/ml) or concanavalin A (1 μg/ml) (Sigma Chemical).

Techniques: Enzyme-linked Immunosorbent Assay

Journal:

Article Title: Prime-Boost Immunization Schedules Based on Influenza Virus and Vaccinia Virus Vectors Potentiate Cellular Immune Responses against Human Immunodeficiency Virus Env Protein Systemically and in the Genitorectal Draining Lymph Nodes

doi: 10.1128/JVI.77.12.7048-7057.2003

Figure Lengend Snippet: Comparison of levels of expression of gp160 between cells infected with VV WR Env and MVA Env. (A) Western blot. 3T3 and BHK-21 cells were infected (5 PFU/cell) with VV WR Env or MVA Env, and at various times postinfection cells were collected and Env expression was analyzed by Western blot with a specific anti-gp160 IIIB antibody. (B) Immunofluorescence analysis under permeable conditions. 3T3 cells were infected (5 PFU/cell) with recombinant VV WR Env, MVA Env, or MVAluc, and at 24 h postinfection cells were fixed, permeabilized, and incubated with polyclonal gp120 IIIB antibody to show Env, with antibody against wheat germ to show the Golgi, or with To-Pro to show the DNA. To the right is the color merging to show colocalization of gp160 and Golgi compartments. (C) Immunofluorescence analysis under nonpermeable conditions. 3T3 cells were infected (1 PFU/cell) with recombinant VV WR Env, MVA Env, or MVAluc, and at 18 h postinfection cells were fixed, nonpermeabilized, and incubated with rabbit polyclonal gp120 IIIB antibody.

Article Snippet: Lymphocytes were cultured in triplicate (10 6 cells/well) in 96-well microtiter flat-bottomed plates and stimulated with purified gp160 protein (Intracel Corporation, Cambridge, Mass.) (1 μg/ml) or concanavalin A (1 μg/ml) (Sigma Chemical).

Techniques: Expressing, Infection, Western Blot, Immunofluorescence, Recombinant, Incubation

Journal:

Article Title: Prime-Boost Immunization Schedules Based on Influenza Virus and Vaccinia Virus Vectors Potentiate Cellular Immune Responses against Human Immunodeficiency Virus Env Protein Systemically and in the Genitorectal Draining Lymph Nodes

doi: 10.1128/JVI.77.12.7048-7057.2003

Figure Lengend Snippet: Pattern of specific cytokine secretion after mucosal immunization. Cell suspensions of spleens (A) or genitorectal lymph nodes (B) of mice in Fig. ​Fig.4,4, obtained 14 days after the booster, were in vitro restimulated with gp160 or RPMI (negative control), and 72 h later cell culture supernatants were harvested and evaluated for IFN-γ and IL-4 by ELISA. Bars represent the mean with standard deviation of triplicates from cell cultures gp160-stimulated; nonspecific cytokine levels found in negative controls were subtracted.

Article Snippet: Lymphocytes were cultured in triplicate (10 6 cells/well) in 96-well microtiter flat-bottomed plates and stimulated with purified gp160 protein (Intracel Corporation, Cambridge, Mass.) (1 μg/ml) or concanavalin A (1 μg/ml) (Sigma Chemical).

Techniques: In Vitro, Negative Control, Cell Culture, Enzyme-linked Immunosorbent Assay, Standard Deviation