Journal: Cells

Article Title: Pharmacological Evidence of the Important Roles of CCR1 and CCR3 and Their Endogenous Ligands CCL2/7/8 in Hypersensitivity Based on a Murine Model of Neuropathic Pain

doi: 10.3390/cells12010098

Figure Lengend Snippet: Effect of a single (20 μg/5 μL) i.t. administration of J113863, SB328437 or UCB35625 on mechanical hypersensitivity ( A , C , E ) in male mice as measured on the 2nd, 12th, and 28th days after chronic constriction injury of the sciatic nerve. Measurements were performed 1, 3, 5 and 24 h after substance injection. The horizontal dotted line shows the cut off value. Moreover, the data obtained were analyzed as areas under the curve ( B , D , F ). The data are presented as the means ± SEMs ( A , C , E ) or total area under curve ± SEMs ( B , D , F ). Intergroup differences were analyzed using one-way ANOVA with Bonferroni’s post hoc test for multiple comparisons. # p < 0.05, ## p < 0.01, and ### p < 0.001 indicate a significant difference compared with the vehicle-treated group. ^ p < 0.05 and ^^ p < 0.001 indicate differences between J11 and SB, § p < 0.05, §§ p < 0.001 indicate a difference between J11 and UCB; ᴏᴏ p < 0.01, ᴏᴏᴏ p < 0.001 indicate a significant difference between SB and UCB. The quantity of animals used in the experiment was as follows: 2nd day: V ( n = 10), J11 ( n = 7–8), SB ( n = 10), UCB ( n = 6); 12th day: V ( n = 7), J11 ( n = 7), SB ( n = 10), UCB ( n = 5); 28th day: V ( n = 6), J11 ( n = 5), SB ( n = 7), UCB ( n = 5). Abbreviations: J11—J113863, SB—SB328437, UCB—UCB35625, V—vehicle.

Article Snippet: J113863, SB328437, UCB35625 and J113863+SB328437 (Tocris) were dissolved in 70% DMSO and administered once i.t. to operated male and female animals on the 12th day and post CCI at a dose of 20 μg/5 μL each.

Techniques: Injection

Journal: Cells

Article Title: Pharmacological Evidence of the Important Roles of CCR1 and CCR3 and Their Endogenous Ligands CCL2/7/8 in Hypersensitivity Based on a Murine Model of Neuropathic Pain

doi: 10.3390/cells12010098

Figure Lengend Snippet: Effect of a single (20 μg/5 μL) i.t . administration of J113863, SB328437 and UCB35625 on thermal hypersensitivity ( A , C , E ) in male mice as measured on the 2nd, 12th, and 28th days after chronic constriction injury of the sciatic nerve. Measurements were performed 1, 3, 5 and 24 h after substance injection. The horizontal dotted line shows the cutoff value. Moreover, the data obtained were analyzed as areas under the curve ( B , D , F ). The data are presented as the means ± SEMs ( A , C , E ) or total area under curve ± SEMs ( B , D , F ). Intergroup differences were analyzed using one-way ANOVA with Bonferroni’s post hoc test for multiple comparisons. # p < 0.05, ## p < 0.01, and ### p < 0.001 indicate a significant difference compared with the vehicle-treated group. ^ p < 0.05, and ^^^ p < 0.001 indicate differences between J11 and SB, § p < 0.05, §§ p < 0.01, and §§§ p < 0.001 indicate differences between J11 and UCB; ᴏᴏ p < 0.01 indicates a significant difference between SB and UCB. The quantity of animals used in the experiment was as follows: 2nd day: V ( n = 10), J11 ( n = 8), SB ( n = 9–10), UCB ( n = 6); 12th day: V ( n = 7), J11 ( n = 7), SB ( n = 8–10), UCB ( n = 5); 28th day: V ( n = 6), J11 ( n = 5), SB ( n = 7), UCB ( n = 5). Abbreviations: J11—J113863, SB—SB328437, UCB—UCB35625, V—vehicle.

Article Snippet: J113863, SB328437, UCB35625 and J113863+SB328437 (Tocris) were dissolved in 70% DMSO and administered once i.t. to operated male and female animals on the 12th day and post CCI at a dose of 20 μg/5 μL each.

Techniques: Injection

Journal: Cells

Article Title: Pharmacological Evidence of the Important Roles of CCR1 and CCR3 and Their Endogenous Ligands CCL2/7/8 in Hypersensitivity Based on a Murine Model of Neuropathic Pain

doi: 10.3390/cells12010098

Figure Lengend Snippet: Effect of a single (20 μg/5 μL) i.t. administration of J113863, SB328437 or UCB35625 on motor coordination in male mice on the 2nd, 12th and 28th days after chronic constriction injury of the sciatic nerve. Measurements in a rotarod apparatus were performed 2 h after single drug administration. The data are presented as the means ± SEMs. Intergroup differences were analyzed using one-way ANOVA with Bonferroni’s post hoc test for multiple comparisons. The quantity of animals used in the experiment was as follows: 2nd day: V ( n = 10), J11 ( n = 7), SB ( n = 10), UCB ( n = 6); 12th day: V ( n = 7), J11 ( n = 8), SB ( n = 10), UCB ( n = 5); 28th day: V ( n = 6), J11 ( n = 5), SB ( n = 7), UCB ( n = 5). Abbreviations: J11—J113863, SB—SB328437, UCB—UCB35625, V—vehicle.

Article Snippet: J113863, SB328437, UCB35625 and J113863+SB328437 (Tocris) were dissolved in 70% DMSO and administered once i.t. to operated male and female animals on the 12th day and post CCI at a dose of 20 μg/5 μL each.

Techniques: Injection

Journal: Cells

Article Title: Pharmacological Evidence of the Important Roles of CCR1 and CCR3 and Their Endogenous Ligands CCL2/7/8 in Hypersensitivity Based on a Murine Model of Neuropathic Pain

doi: 10.3390/cells12010098

Figure Lengend Snippet: Effect of a single (20 μg/5 μL) i.t. administration of J113863, SB328437, UCB35625 or J11386 + SB328437 (20 μg/5 μL + 20 μg/5) on % of the maximal possible effect on the 12th day after chronic constriction injury of the sciatic nerve in male and female mice. Measurements in a von Frey and cold plate apparatus were performed 3 h after single drug administration. The data are presented as the means ± SEMs. Intergroup differences were analyzed using one-way ANOVA with Bonferroni’s post hoc test for multiple comparisons. t test was used to evaluate sex differences in each substance. # p < 0.05, ## p < 0.01 and ### p < 0.001 indicate a significant difference compared with the vehicle group; §§ p < 0.01 indicates a significant difference between male and female. The quantity of animals used in the experiment was as follows: male mice: (von Frey: V ( n = 10), J11 ( n = 7), SB ( n = 10), UCB ( n = 5), J11 + SB ( n = 8)), (cold plate: V ( n = 7), J11 ( n = 8), SB ( n = 8), UCB ( n = 5); J11 + SB ( n = 8)); female mice: (von Frey: V ( n = 8), J11 ( n = 6), SB ( n = 8), UCB ( n = 6), J11 + SB ( n = 6), (cold plate: V ( n = 8), J11 ( n = 6), SB ( n = 8), UCB ( n = 6); J11 + SB ( n = 6)). Abbreviations: J11—J113863, SB—SB328437, UCB—UCB35625, V—vehicle.

Article Snippet: J113863, SB328437, UCB35625 and J113863+SB328437 (Tocris) were dissolved in 70% DMSO and administered once i.t. to operated male and female animals on the 12th day and post CCI at a dose of 20 μg/5 μL each.

Techniques:

Journal: Cells

Article Title: Pharmacological Evidence of the Important Roles of CCR1 and CCR3 and Their Endogenous Ligands CCL2/7/8 in Hypersensitivity Based on a Murine Model of Neuropathic Pain

doi: 10.3390/cells12010098

Figure Lengend Snippet: Selective inhibition of CCL2/7/8 production by bindarit, as well as blocking of CCR1 and CCR3 by selective (J113863, SB328437) or dual (UCB35625) antagonists, reduced even fully developed mechanical and thermal hypersensitivity in murine model of neuropathic pain.

Article Snippet: J113863, SB328437, UCB35625 and J113863+SB328437 (Tocris) were dissolved in 70% DMSO and administered once i.t. to operated male and female animals on the 12th day and post CCI at a dose of 20 μg/5 μL each.

Techniques: Inhibition, Blocking Assay

Journal: Cells

Article Title: Pharmacological Evidence of the Important Roles of CCR1 and CCR3 and Their Endogenous Ligands CCL2/7/8 in Hypersensitivity Based on a Murine Model of Neuropathic Pain

doi: 10.3390/cells12010098

Figure Lengend Snippet: Effect of a single (20 μg/5 μL) i.t. administration of J113863, SB328437 or UCB35625 on mechanical hypersensitivity ( A , C , E ) in male mice as measured on the 2nd, 12th, and 28th days after chronic constriction injury of the sciatic nerve. Measurements were performed 1, 3, 5 and 24 h after substance injection. The horizontal dotted line shows the cut off value. Moreover, the data obtained were analyzed as areas under the curve ( B , D , F ). The data are presented as the means ± SEMs ( A , C , E ) or total area under curve ± SEMs ( B , D , F ). Intergroup differences were analyzed using one-way ANOVA with Bonferroni’s post hoc test for multiple comparisons. # p < 0.05, ## p < 0.01, and ### p < 0.001 indicate a significant difference compared with the vehicle-treated group. ^ p < 0.05 and ^^ p < 0.001 indicate differences between J11 and SB, § p < 0.05, §§ p < 0.001 indicate a difference between J11 and UCB; ᴏᴏ p < 0.01, ᴏᴏᴏ p < 0.001 indicate a significant difference between SB and UCB. The quantity of animals used in the experiment was as follows: 2nd day: V ( n = 10), J11 ( n = 7–8), SB ( n = 10), UCB ( n = 6); 12th day: V ( n = 7), J11 ( n = 7), SB ( n = 10), UCB ( n = 5); 28th day: V ( n = 6), J11 ( n = 5), SB ( n = 7), UCB ( n = 5). Abbreviations: J11—J113863, SB—SB328437, UCB—UCB35625, V—vehicle.

Article Snippet: J113863 (CCR1 antagonist), SB328437 (CCR3 antagonist), and UCB35625 (CCR1/CCR3 antagonist) (all obtained from Tocris, Bristol, UK) were dissolved in 70% DMSO and administered once i.t. to operated male animals on the 2nd, 12th and 28th days post CCI at a dose of 20 μg/5 μL.

Techniques: Injection

Journal: Cells

Article Title: Pharmacological Evidence of the Important Roles of CCR1 and CCR3 and Their Endogenous Ligands CCL2/7/8 in Hypersensitivity Based on a Murine Model of Neuropathic Pain

doi: 10.3390/cells12010098

Figure Lengend Snippet: Effect of a single (20 μg/5 μL) i.t . administration of J113863, SB328437 and UCB35625 on thermal hypersensitivity ( A , C , E ) in male mice as measured on the 2nd, 12th, and 28th days after chronic constriction injury of the sciatic nerve. Measurements were performed 1, 3, 5 and 24 h after substance injection. The horizontal dotted line shows the cutoff value. Moreover, the data obtained were analyzed as areas under the curve ( B , D , F ). The data are presented as the means ± SEMs ( A , C , E ) or total area under curve ± SEMs ( B , D , F ). Intergroup differences were analyzed using one-way ANOVA with Bonferroni’s post hoc test for multiple comparisons. # p < 0.05, ## p < 0.01, and ### p < 0.001 indicate a significant difference compared with the vehicle-treated group. ^ p < 0.05, and ^^^ p < 0.001 indicate differences between J11 and SB, § p < 0.05, §§ p < 0.01, and §§§ p < 0.001 indicate differences between J11 and UCB; ᴏᴏ p < 0.01 indicates a significant difference between SB and UCB. The quantity of animals used in the experiment was as follows: 2nd day: V ( n = 10), J11 ( n = 8), SB ( n = 9–10), UCB ( n = 6); 12th day: V ( n = 7), J11 ( n = 7), SB ( n = 8–10), UCB ( n = 5); 28th day: V ( n = 6), J11 ( n = 5), SB ( n = 7), UCB ( n = 5). Abbreviations: J11—J113863, SB—SB328437, UCB—UCB35625, V—vehicle.

Article Snippet: J113863 (CCR1 antagonist), SB328437 (CCR3 antagonist), and UCB35625 (CCR1/CCR3 antagonist) (all obtained from Tocris, Bristol, UK) were dissolved in 70% DMSO and administered once i.t. to operated male animals on the 2nd, 12th and 28th days post CCI at a dose of 20 μg/5 μL.

Techniques: Injection

Journal: Cells

Article Title: Pharmacological Evidence of the Important Roles of CCR1 and CCR3 and Their Endogenous Ligands CCL2/7/8 in Hypersensitivity Based on a Murine Model of Neuropathic Pain

doi: 10.3390/cells12010098

Figure Lengend Snippet: Effect of a single (20 μg/5 μL) i.t. administration of J113863, SB328437 or UCB35625 on motor coordination in male mice on the 2nd, 12th and 28th days after chronic constriction injury of the sciatic nerve. Measurements in a rotarod apparatus were performed 2 h after single drug administration. The data are presented as the means ± SEMs. Intergroup differences were analyzed using one-way ANOVA with Bonferroni’s post hoc test for multiple comparisons. The quantity of animals used in the experiment was as follows: 2nd day: V ( n = 10), J11 ( n = 7), SB ( n = 10), UCB ( n = 6); 12th day: V ( n = 7), J11 ( n = 8), SB ( n = 10), UCB ( n = 5); 28th day: V ( n = 6), J11 ( n = 5), SB ( n = 7), UCB ( n = 5). Abbreviations: J11—J113863, SB—SB328437, UCB—UCB35625, V—vehicle.

Article Snippet: J113863 (CCR1 antagonist), SB328437 (CCR3 antagonist), and UCB35625 (CCR1/CCR3 antagonist) (all obtained from Tocris, Bristol, UK) were dissolved in 70% DMSO and administered once i.t. to operated male animals on the 2nd, 12th and 28th days post CCI at a dose of 20 μg/5 μL.

Techniques: Injection

Journal: Cells

Article Title: Pharmacological Evidence of the Important Roles of CCR1 and CCR3 and Their Endogenous Ligands CCL2/7/8 in Hypersensitivity Based on a Murine Model of Neuropathic Pain

doi: 10.3390/cells12010098

Figure Lengend Snippet: Effect of a single (20 μg/5 μL) i.t. administration of J113863, SB328437, UCB35625 or J11386 + SB328437 (20 μg/5 μL + 20 μg/5) on % of the maximal possible effect on the 12th day after chronic constriction injury of the sciatic nerve in male and female mice. Measurements in a von Frey and cold plate apparatus were performed 3 h after single drug administration. The data are presented as the means ± SEMs. Intergroup differences were analyzed using one-way ANOVA with Bonferroni’s post hoc test for multiple comparisons. t test was used to evaluate sex differences in each substance. # p < 0.05, ## p < 0.01 and ### p < 0.001 indicate a significant difference compared with the vehicle group; §§ p < 0.01 indicates a significant difference between male and female. The quantity of animals used in the experiment was as follows: male mice: (von Frey: V ( n = 10), J11 ( n = 7), SB ( n = 10), UCB ( n = 5), J11 + SB ( n = 8)), (cold plate: V ( n = 7), J11 ( n = 8), SB ( n = 8), UCB ( n = 5); J11 + SB ( n = 8)); female mice: (von Frey: V ( n = 8), J11 ( n = 6), SB ( n = 8), UCB ( n = 6), J11 + SB ( n = 6), (cold plate: V ( n = 8), J11 ( n = 6), SB ( n = 8), UCB ( n = 6); J11 + SB ( n = 6)). Abbreviations: J11—J113863, SB—SB328437, UCB—UCB35625, V—vehicle.

Article Snippet: J113863 (CCR1 antagonist), SB328437 (CCR3 antagonist), and UCB35625 (CCR1/CCR3 antagonist) (all obtained from Tocris, Bristol, UK) were dissolved in 70% DMSO and administered once i.t. to operated male animals on the 2nd, 12th and 28th days post CCI at a dose of 20 μg/5 μL.

Techniques:

Journal: Cells

Article Title: Pharmacological Evidence of the Important Roles of CCR1 and CCR3 and Their Endogenous Ligands CCL2/7/8 in Hypersensitivity Based on a Murine Model of Neuropathic Pain

doi: 10.3390/cells12010098

Figure Lengend Snippet: Selective inhibition of CCL2/7/8 production by bindarit, as well as blocking of CCR1 and CCR3 by selective (J113863, SB328437) or dual (UCB35625) antagonists, reduced even fully developed mechanical and thermal hypersensitivity in murine model of neuropathic pain.

Article Snippet: J113863 (CCR1 antagonist), SB328437 (CCR3 antagonist), and UCB35625 (CCR1/CCR3 antagonist) (all obtained from Tocris, Bristol, UK) were dissolved in 70% DMSO and administered once i.t. to operated male animals on the 2nd, 12th and 28th days post CCI at a dose of 20 μg/5 μL.

Techniques: Inhibition, Blocking Assay

Journal: bioRxiv

Article Title: Memory CD8 + T cells mediate early pathogen-specific protection through localized delivery of chemokines and IFNγ to clusters of inflammatory monocytes

doi: 10.1101/2021.03.01.433468

Figure Lengend Snippet: WT mice transferred with OT-I cells were immunized with VSV- Ova (A) or Lm- Ova (B, C) and challenged or not, 6 wks later with Lm or Lm- Ova. Spleens from 16 hr-challenged or unchallenged mice were harvested, and cells incubated for 4-6 hrs with brefeldin A prior to staining for expression of CD11b, Ly6C and Ly6G cell surface markers and (A) indicated intracellular effector and chemotactic markers or (B) expression of CCR1, CCR5 and XCR1 chemotactic receptors. In (C), monocytes and neutrophils from the spleens of challenged mice were flow-sorted and either stimulated or not with indicated recombinant chemokines or HK Lm , with or without CCR5 and CCR1 inhibitors, prior to staining for intracellular TNFα. (D) BM from WT (CD45.1 +/- ) and Ifngr1 -/- (CD45.2 +/+ ) mice were co-transferred to CD45.1 +/+ recipient mice immunized with Lm -Ova 6 weeks before, and immediately challenged with Lm . 16 hrs later, spleen cells were stained for expression of CCR5 and XCR1 on monocytes. (E) Lm- Ova immunized mice were challenged with Lm -Ova or not, and 1 hr before sacrifice, 5 μg Ly6C-PE mAb was injected i.v. to the hosts. Spleens were harvested and cells stained for cell surface CD11b, Ly6C-PerCpCy5.5, ICAM-1, CD86 and intracellular TNFα and CXCL9. After gating on Ly6C-PerCpCy5.5 + monocytes, Ly6C-PE hi and Ly6C-PE low monocytes were identified and further analyzed for indicated marker expression. Representative FACS dot plots are shown and bar graphs pool 2 independent replicate experiments with n=6 (A, D) and 5 (B, C and E) mice. P- values are indicated.

Article Snippet: In CCR5 and CCR1 blocking experiments, cells were incubated with both CCR5 (1μM, Maraviroc, Cayman Chemical Company) and CCR1 (1μM, J113863, Santa Cruz) chemical inhibitors or control DMSO 30 min prior to adding recombinant chemokines or HK Lm .

Techniques: Incubation, Staining, Expressing, Recombinant, Injection, Marker

Journal: bioRxiv

Article Title: Memory CD8 + T cells mediate early pathogen-specific protection through localized delivery of chemokines and IFNγ to clusters of inflammatory monocytes

doi: 10.1101/2021.03.01.433468

Figure Lengend Snippet: (A, B) Mice grafted with OT-I cells were immunized with Lm - Ova and 6 wks later challenged with 10 6 Lm- Ova. Sixteen hrs post-challenge, monocytes and neutrophils were sorted from spleen and stimulated for 4 hrs with or without recombinant chemokines at indicated concentrations, or with HK Lm and with or without CCR1/5 inhibitors (depicted in ). Cells were next stained for cell surface expression of CD11b, Ly6C, Ly6G and intracellular TNFα. Representative FACS dot plots are shown. (B) Graphs show TNFα + monocytes and neutrophils frequency after 4hrs incubation with recombinant chemokines (1 and 0.3µg) or HK Lm . Bar graphs (each symbol is 1 mouse) represent the pool of 2 independent replicate experiments with p-values indicated. (C) Splenocytes from WT naive mice injected with PBS were stimulated in vitro with HK Lm or rCCL3 (1µg). Cells were stained as above. Representative FACS dot plots with summary bar graphs (each symbol is 1 mouse) with indicated p-value are shown. (D) TNFα expression in flow-sorted neutrophils as depicted in , incubated with CCR5 and CCR1 inhibitors or DMSO vehicle. Bar graphs (each symbol is 1 mouse) represent the pool of 2 independent replicate experiments with p-values indicated.

Article Snippet: In CCR5 and CCR1 blocking experiments, cells were incubated with both CCR5 (1μM, Maraviroc, Cayman Chemical Company) and CCR1 (1μM, J113863, Santa Cruz) chemical inhibitors or control DMSO 30 min prior to adding recombinant chemokines or HK Lm .

Techniques: Recombinant, Staining, Expressing, Incubation, Injection, In Vitro

Journal: Biomedicines

Article Title: Analysing the Combined Effects of Radiotherapy and Chemokine Receptor 5 Antagonism: Complementary Approaches to Promote T Cell Function and Migration in Oesophageal Adenocarcinoma

doi: 10.3390/biomedicines12040819

Figure Lengend Snippet: Expanded circulating T cells from OAC patients expressed higher levels of CCR5, which was further upregulated on the surface of CD8 + T cells by clinically relevant doses of irradiation. ( a ) PBMCs isolated from treatment−naïve OAC donors ( n = 7) and age—matched HDs ( n = 6) were activated with plate−bound anti−CD3 and anti−CD28 agonists for 72 h, receiving 2 × 1.8 Gy fractions of irradiation (irradiated, IR) on day 1 and day 2, 24 h apart, or were non−irradiated, NIR. The frequencies of T cells in HDs and CDs expressing CCR1 ( b ), CCR5 ( c , d ), and CX 3 CR1 ( e ) were assessed by flow cytometry. The effect of IR on CCR1, CCR5, and CX 3 CR1 expression on T cells from HDs ( f ) and CDs ( g ) was also assessed by flow cytometry and depicted in heat maps shown (% cells expressing). The effect of IR on CCR5 expression on T cells from CDs is displayed in ( h , i ). All analyses were conducted on viable T cells using a zombie dye to exclude dead cells, and fluorescence minus-one controls (FMO) were used for gating analysis. Mann−Whitney tests were used to compare between HD and CD groups. Wilcoxon signed-rank tests were used to compare the effect of NIR with IR in the same donors. * p < 0.05, ** p < 0.01.

Article Snippet: For experiments that included treatment with antagonists, in the last 24 h of the T cell activation process, PBMCs were treated with 1 nM CCR1 antagonist J113863 (Axon MedChem, Reston, VA, USA), 80 μM of CCR5 antagonist Maraviroc (Axon MedChem, Reston, VA, USA), 245 nM CX 3 CR1 antagonist AZD8798 (Axon MedChem, Reston, VA, USA), or vehicle control (0.01% DMSO).

Techniques: Irradiation, Isolation, Expressing, Flow Cytometry, Fluorescence, MANN-WHITNEY

Journal: Biomedicines

Article Title: Analysing the Combined Effects of Radiotherapy and Chemokine Receptor 5 Antagonism: Complementary Approaches to Promote T Cell Function and Migration in Oesophageal Adenocarcinoma

doi: 10.3390/biomedicines12040819

Figure Lengend Snippet: CCR5 antagonism significantly increased IFN-γ production by OAC patient−derived CD4 + T helper cells. ( a ) PBMCs isolated from treatment-naïve OAC donors ( n = 5) and age−matched non-cancer donors ( n = 6) were activated with anti-CD3 and anti-CD28 agonists for 72 h and treated with J113863 (CCR1 antagonist) and RANTES, AZD8987 (CX 3 CR1 antagonist) and fractalkine or Maraviroc (CCR5 antagonist) and MIP-1α. The PBMCs also received 2 × 1.8 Gy fractions of irradiation on day 1 and day 2, 24 h apart, or were non-irradiated (NIR). The frequency of cells producing IFN-γ and CD107a was assessed by intracellular and extracellular flow cytometry, respectively. ( b – d ) heat maps depicting the effect of activating or antagonising the CCR1−RANTES, CCR5-MIP-1α, and CX 3 CR1−fractalkine pathways on the percentage of CD4 + and CD8 + T cells producing IFN-γ, respectively, relative to the untreated control. ( e ) Graphical display showing the effect of CCR5 antagonism on the production of IFN-γ by T cells, with representative dot plots shown in ( f ). ( g ) The effect of activating or antagonizing CCR1-RANTES, CCR5-MIP-1α, and CX 3 CR1-fractalkine pathway on CD107a degranulation by CD8 + T cells. All analyses were conducted on viable T cells using a zombie dye to exclude dead cells, and FMO controls were used for gating analysis. Paired parametric t -test was used to compare between two groups * p < 0.05.

Article Snippet: For experiments that included treatment with antagonists, in the last 24 h of the T cell activation process, PBMCs were treated with 1 nM CCR1 antagonist J113863 (Axon MedChem, Reston, VA, USA), 80 μM of CCR5 antagonist Maraviroc (Axon MedChem, Reston, VA, USA), 245 nM CX 3 CR1 antagonist AZD8798 (Axon MedChem, Reston, VA, USA), or vehicle control (0.01% DMSO).

Techniques: Derivative Assay, Isolation, Irradiation, Flow Cytometry, Control