mouse mab α cd8  (Thermo Fisher)

Journal: Immunity

Article Title: Targeted complement inhibition at synapses prevents microglial synaptic engulfment and synapse loss in demyelinating disease

doi: 10.1016/j.immuni.2019.12.004

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: The following primary monoclonal (mAb) and polyclonal (pAb) antibodies have been used: mouse mAb α-ALDH1L1 (clone N103/39, Millipore, MABN495, 1:1000), mouse mAb α-APP (clone 22C11, Millipore, MAB348, 1:200), rabbit mAb α-C1q (clone 4.8, Abcam, ab182451, 1:100), rat mAb α-C3 (clone 11H-9, Abcam, ab11862, 1:500), rabbit pAb α-CASPR (provided by Matthew N. Rasband, 1:100), rabbit mAb α-CD3 (clone SP7, Abcam, ab16669, 1:100), mouse mAb α-CD8 (clone 8/114B, Thermo Fisher Scientific, MA5–13473, 1:20), rat mAb α-CD45 (clone IBL-3/16, BioRad, MCA1388, 1:100), mouse mAb α-CD68 (clone KP1, Abcam, ab955, 1:200), rat mAb α-CD68 (clone FA-11, AbD Serotec, MCA1957, 1:1000), rat mAb α-Clec7a (InvivoGen, mabg-mdect, 1:200), rabbit pAb α-Clec12a (LSBio, LS-C377776, 1:500), rabbit pAb α-cleaved caspase-3 (Cell Signaling Technologies, #9661, 1:200), mouse mAb α-Crry (clone TLD-1C11, Santa Cruz, sc-53530, 1:100), chicken pAb α-EGFP (Abcam, ab13970, 1:500), mouse mAb α-FoxP3 (clone 206D, BioLegend, 320102, 1:50), mouse mAb α-GFAP (clone G-A-5, Sigma, G3893, 1:500), rabbit pAb α-Homer1 (Synaptic Systems, #160003, 1:1000), rabbit pAb α-Iba1 (Wako Chemicals, #019–19741, 1:500), rat mAb α-LAMP2 (clone GL2A7, Abcam, ab13524, 1:200), rat mAb α-Ly6C (clone HK1.4, BioLegend, 128016, 1:300), mouse mAb α-MAG (clone 513, Millipore, MAB1567, 1:100), rat mAb α-MBP (clone 12, Millipore, MAB386, 1:500), mouse mAb α-MOG (clone 8–18C5, Millipore, MAB5680, 1:200), rabbit pAb α-Neurofilament 200 (Sigma, N4142, 1:1000), chicken pAb α-NeuN (Millipore, ABN91, 1:1000), rabbit pAb α-P2RY12 (AnaSpec, AS-55043A, 1:2000), rat mAb α-P2RY12 (clone S16007D, BioLegend, 848002, 1:100), mouse mAb α-PSD-95 (clone 6G6–1C9, Millipore, MAB1596, 1:100), guinea pig pAb α-RBPMS (PhosphoSolutions, 1832-RBPMS, 1:500), rabbit pAb α-βIV-spectrin (provided by Matthew N. Rasband, 1:100), guinea pig pAb α-VGluT1 (Millipore, ab5905, 1:2000), and guinea pig pAb α-VGluT2 (Millipore, ab2251, 1:2000).

Techniques: Virus, Recombinant, Adjuvant, Electron Microscopy, Cloning, Plasmid Preparation, RNAscope, Multiplex Assay, In Situ, Sequencing, Generated, Software, Imaging

Journal: Nature Communications

Article Title: An mRNA vaccine against monkeypox virus inhibits infection by co-activation of humoral and cellular immune responses

doi: 10.1038/s41467-025-58328-x

Figure Lengend Snippet: a Schematic diagram of the immunization and challenge experiments for multicomponent mRNA vaccines. Created in BioRender. Tian, C. (2025) https://BioRender.com/s30e801 . The designed 4-valent, 8-valent, and 12-valent mRNA vaccines, comprising 4, 8, and 12 antigens, respectively. These vaccines were formulated by an equal-ratio mixture of mRNAs for each component and named as Mix-4, Mix-8, and Mix-12, accordingly. b Neutralizing antibody titers (NT 50 ) induced by different mRNA vaccines. The neutralizing antibodies titers (NT 50 ) induced by Mix-4, Mix-8, and Mix-12 were measured by plaque reduction neutralization tests. PBS-immunized groups were included as controls. NT 50 is indicated as mean ± SEM. ( n = 6 per group). c Heatmaps demonstrating IgG subclass levels in the serum from immunized mice ( n = 6 per group). Heatmaps were created using the hiplot online platform ( https://hiplot.com.cn/ ). d Protective potency of plasma passive transfusion. Three-week-old BALB/c mice ( n = 6) were transfused with 200 μl of plasma from immunized mice and challenged with MPXV (1.0 × 10 6 PFU) via intranasal inoculation. Viral titers in the lungs were measured using a standard plaque assay. e Proliferation of epitope-specific splenocytes producing IFN-γ following stimulation. IFN-γ-producing epitope-specific splenocytes were assessed by ELISpot. Each point represents the mean of 3 technical replicates, with a limit of detection (LOD) of 10, and representative spots are shown. f Protective potency of CD8 + T cells and CD4 + T cells transfusion. (Created in BioRender. Tian, C. (2025) https://BioRender.com/h28r862 ). Three-week-old BALB/c mice ( n = 6) were transfused with 2.0 × 10 7 CD8 + or CD4 + T cells sorted from immunized mice and intranasally inoculated with MPXV at the dose of 1.0 × 10 6 PFU. Mice were sacrificed 4 days post-infection, and lungs were harvested for viral load detection. Data are shown as mean ± SEM from individual mice ( b , d , e , f ). Two independent experiments were performed with 2 technical replicates ( b – f ). P values were determined by one-way ANOVA with Tukey’s multiple comparison post-hoc test. Source data are provided as a file.

Article Snippet: MPX - EPs (3 μg) or PBS immunized HLA-A*02:01/DR1 transgenic mice ( n = 6 under animal immunization c.) were injected intraperitoneally with an anti-mouse anti-CD8-α (IgG2b) mAb (Bio X Cell, Cat. No# BE0061, 100 μg/mouse) or IgG2b isotype control mAb (Bio X Cell, Cat. No# BE0090, 100 μg/mouse) on Days-1 and 0 of the challenge.

Techniques: Vaccines, Neutralization, Clinical Proteomics, Plaque Assay, Enzyme-linked Immunospot, Infection, Comparison

Journal: Nature Communications

Article Title: An mRNA vaccine against monkeypox virus inhibits infection by co-activation of humoral and cellular immune responses

doi: 10.1038/s41467-025-58328-x

Figure Lengend Snippet: a Schematic diagram of the MPX-EPs mRNA design and evaluation. Created in BioRender. Tian, C. (2025) https://BioRender.com/b59j557 . Predicted CTL epitopes enriched peptides were linked with A-A-Y motifs, with a sequence encoding ubiquitin (Ub) added to the N-terminus of the peptides. b – e Evaluation of the cellular immune response. The percentages of the lymphocyte subsets of CD45 + CD4 + T cells ( b , left ) and CD45 + CD8 + T cells ( b , right ) in the spleens of immunized HLA-A*02:01/DR1 transgenic mice were assessed by flow cytometry. The proliferation of MPX-EPs-specific, IFN-γ producing splenocytes was measured by ELISpot ( c ). Each point represents the mean of 3 technical replicates, with a lower limit of detection (LLOD) of 5, and representative spots were shown ( c ). The percentage of TNF-α and IFN-γ positive CD8 + T cells was measured ( d , e ). Representative flow cytometry plots are presented, with symbols representing the means of two technical replicates per individual mouse. f – h MPX-EPs-induced protection against MPXV. HLA-A*02:01/DR1 transgenic mice were intramuscularly vaccinated with two doses of MPX-EPs (3 μg/dose). At 20 days post-booster immunization, the immunized mice were injected (i.p.) with an anti-CD8-α antibody (to deplete CD8 + T cells) or an IgG2b isotype control antibody (without CD8 + T-cells depletion) (200 mg/mouse) twice with a 24 h interval. The mice were then infected with the MPXV (1.0 × 10 7 PFU/mouse). Viral titers after 4 days post-inoculation in lung tissues from HLA-A*02:01/DR1 transgenic mice, with and without CD8 + T cell depletion ( n = 6), were determined by a plaque assay with two technical replicates ( f ). The LLOD was 20 PFU per gram of tissue. Lung tissue pathology was evaluated by HE staining ( g ) and pathological scoring ( h ), Scale bar is 200 μm. Statistical significance was calculated via two-sided unpaired t-test ( b , c , e ) or one-way ANOVA with Tukey’s multiple comparisons post-hoc tests ( f , h ). Experiments were repeated twice independently, and similar results were obtained. Both independent experiments contain 3 technical repeats ( b – h ). Data are presented as mean ± SEM ( b , c, e , f , h ). Source data are provided as a file.

Article Snippet: MPX - EPs (3 μg) or PBS immunized HLA-A*02:01/DR1 transgenic mice ( n = 6 under animal immunization c.) were injected intraperitoneally with an anti-mouse anti-CD8-α (IgG2b) mAb (Bio X Cell, Cat. No# BE0061, 100 μg/mouse) or IgG2b isotype control mAb (Bio X Cell, Cat. No# BE0090, 100 μg/mouse) on Days-1 and 0 of the challenge.

Techniques: Sequencing, Ubiquitin Proteomics, Transgenic Assay, Flow Cytometry, Enzyme-linked Immunospot, Injection, Control, Infection, Plaque Assay, Staining

Journal: Nature Communications

Article Title: An mRNA vaccine against monkeypox virus inhibits infection by co-activation of humoral and cellular immune responses

doi: 10.1038/s41467-025-58328-x

Figure Lengend Snippet: a Schematic diagram of the dual immunization design using Mix-12 and MPX-EPs antigens. HLA-A*02:01/DR transgenic mice were immunized with MPX-m-Mix (containing 3 μg MPX-EPs and 12 μg Mix-12 per mouse), MPX-p-Mix (containing 12 μg proteins from Mix-12 and 3 μg peptides from MPX-EPs per mouse), or PBS with 8.7% sucrose as a control. A booster vaccination was administered 3 weeks later. Blood and lymph nodes were collected 1-week post-booster for humoral immune response analysis, while cells from lungs and spleens were collected 3 weeks post-booster for cellular immune response evaluation. Immunized mice ( n = 5 per group) were challenged with MPXV virus (1.0 × 10 7 PFU) 3 weeks after the booster vaccination. b , c Antibody production following immunization against MPXV. MPXV-specific IgG antibodies in the sera collected 21 days after the booster vaccination were detected with ELISA. Neutralizing antibody titers (NT 50 ) were determined by plaque reduction neutralization test. d – f Th1-biased cellular immune response induced by MPX-Mix. Splenocytes from immunized HLA-A*02:01/DR1 transgenic mice ( n = 5) collected 21 days post-booster immunization were re-stimulated ex vivo and subjected to IFN-γ ELISpot ( d ) (lower limit of detection (LLOD) = 20) and IL-4 ELISpot ( e ) (LLOD = 2), with the correlation of IL-4- and IFN-γ-secreting cells shown in the scatter plot ( f ). g – h Analysis of lung-resident CD8 + T cells from immunized mice ( n = 5 per group). Lung cells were stained with anti-CD45-Alexa Fluor™ 700, CD8-PerCP-Cyanine5.5, CD69-FITC, and CD103-AF594. The frequencies of CD45 + CD8 + CD69 + CD103 + T cells were measured by flow cytometry, with the representative flow cytometry plots presented. i – k Protection conferred by MPX-m-Mix or MPX-p-Mix against MPXV. Viral loads in the lungs ( i ) and lung pathology ( j , k ) were evaluated at 4 days post MPXV inoculation (i.n., 1.0 × 10 7 PFU, n = 5 mice per group), Scale bar is 200 μm. Data are representative of two independent experiments with three technical replicates ( b – k ). Statistical significance was determined by one-way ANOVA with Tukey’s multiple comparisons ( n = 5). Data are presented as mean ± SEM ( b – e , h , I , k ). Source data are provided as a file.

Article Snippet: MPX - EPs (3 μg) or PBS immunized HLA-A*02:01/DR1 transgenic mice ( n = 6 under animal immunization c.) were injected intraperitoneally with an anti-mouse anti-CD8-α (IgG2b) mAb (Bio X Cell, Cat. No# BE0061, 100 μg/mouse) or IgG2b isotype control mAb (Bio X Cell, Cat. No# BE0090, 100 μg/mouse) on Days-1 and 0 of the challenge.

Techniques: Transgenic Assay, Control, Virus, Enzyme-linked Immunosorbent Assay, Plaque Reduction Neutralization Test, Ex Vivo, Enzyme-linked Immunospot, Staining, Flow Cytometry

Journal: Nature Communications

Article Title: An mRNA-based T-cell-inducing antigen strengthens COVID-19 vaccine against SARS-CoV-2 variants

doi: 10.1038/s41467-023-38751-8

Figure Lengend Snippet: a Schematic diagram of experimental design. The epitopes for 78 HLA-I alleles were predicted by both the NetMHCpan and IEDB bioinformatic tools. The peptides enriching HLA-I epitopes were ectopically expressed to assess the capability to activate CD8 + T lymphocytes from convalescent COVID-19 patients through a reporter cell-based epitope expression system. b , c Identification of the fragments enriching human cytotoxic T-lymphocyte epitopes in the SARS-CoV-2 proteome. The distribution of potential cytotoxic HLA-I epitopes was predicted by in silico analyses of NetMHCpan ( b ) and IEDB ( c ). Four regions, NSP-3 1443–1605 , NSP-4 232–444 , NSP-6 1–201 , and M 1–113 , with more than 20 effective epitopes (affinity concentration >10 nM) per 100 amino acids were selected as HLA-I epitope-enriched peptides for further investigation, and the three regions, NSP-1 1–180 , NSP-3 1066-1278 , and NSP-14 330–490 with fewest predicted effective epitopes were set as controls. The N protein was included in further investigation. The horizontal lines denote the threshold for selecting the HLA-I epitope enrichment regions. d , e Activation of CD8 + T lymphocytes from convalescent COVID-19 patients. HEK293T cells were transfected to stably express the pHAGE_EF1a_ICADCR, pHAGE_EF1a_IFPGZB, and pHAGE_EF1aHLA vectors. Either HLA-A*02:01 ( d ) or HLA-A*11:01 ( e ) overexpressing HEK293T cells were transfected with the genes encoding the NSP-1 1–180 , NSP-3 1066–1278 , NSP-1 4330–490 , N Full-length , M 1–113 , NSP-3 1443–1605 , NSP-4 232–444 , or NSP-6 1–201 , respectively. At 36 h post-transfection, 1 × 10 6 HEK293T cells were co-cultured with 2 × 10 5 HLA genotype-matched memory CD8 + T cells from convalescent COVID-19 patients ( n = 10 for HLA-A*02:01, and n = 9 for HLA-A*11:01) for 12 h, and activation of CD8 + T cells was characterized by the relative IFP signal intensity in the co-cultred HEK293T cells using the flow cytometric measurement. All data are presented as the mean ± S.E.M from the indicated number of biological repeats. Source data are provided as a Source Data file. Experiments were repeated twice independently with similar results. Both independent experiments contain two technical repeats ( d , e ). Adjusted p values were determined by one-way ANOVA with Tukey’s multiple comparison post-hoc two-sided tests.

Article Snippet: LNP- HLA-EPs (0.5 μg and 10 μg per dose) or LNP -NC (10 μg) immunized HLA-A*02:01/DR1 transgenic mice or HLA-A*11:01/DR1 transgenic mice were injected intraperitoneally with an anti-mouse anti-CD8-α (IgG2b) mAb (Bio X Cell, Cat. No# BE0061, 100 μg/mouse, n = 5) or IgG2b isotype control mAb (Bio X Cell, Cat. No# BE0090, 100 μg/mouse, n = 5) on Days −1 and 0 of the challenge.

Techniques: Expressing, In Silico, Concentration Assay, Activation Assay, Transfection, Stable Transfection, Cell Culture, Comparison

Journal: Nature Communications

Article Title: An mRNA-based T-cell-inducing antigen strengthens COVID-19 vaccine against SARS-CoV-2 variants

doi: 10.1038/s41467-023-38751-8

Figure Lengend Snippet: a Schematic diagram of the HLA-EPs mRNA construct. b , c The expression of HLA-EPs in HEK293T cells. The results are shown by the mean fluorescent intensity (MFI) of HLA-EPs positive cells ( b ) and the representative flow cytometry plots ( c ) of 3 biological replicates containing 3 technical replicates. d–f The proliferation of epitope-specific splenocytes producing IFN-γ following stimulation with the peptide pool. The proliferation of epitope-specific, IFN-γ producing splenocytes was measured by ELISpot. Each point represents the mean of 3 technical replicates, with a limit of detection (LOD) = 2, and representative dots are shown ( d ). The frequency of IFN-γ positive CD8 + T cells was measured by an ICS ( e , f ). The representative flow cytometry plots are presented, and symbols represent individual mice. g – j The CD8 + T-cell-dependent protection of LNP- HLA-EPs against SARS-CoV-2. Both vaccinated HLA-A*02:01/DR1 and HLA-A*11:01/DR1 transgenic mice with or without CD8 + T-cell depletion, were infected with the SARS-CoV-2 B.1.351 variant (1.4 × 10 5 PFU/mouse). Viral titers in lung tissues of HLA-A*02:01/DR1 and HLA-A*11:01/DR1 transgenic mice, with and without CD8 + T-cell depletion after 4 days post-inoculation ( n = 5), were determined by a plaque assay with two technical replicates ( g , h ). The LOD was 10 PFU per gram of tissue. Histopathological changes in the lungs of challenged mice were evaluated by an H&E staining ( i , j ). Images derived from one representative animal in each group with sites of inflammatory cell infiltration (black arrows), blood clots (blue arrows), and alveolar deformation (red arrows) are presented. Scale bar is 50 μm. All data ( b , d – j ) are presented as the mean ± S.E.M. from the indicated number of biological repeats, and statistical significance was calculated via one-way ANOVA with Tukey’s multiple comparison post hoc two-sided tests ( b , d – f , i , j ) or two-way ANOVA with Bonferroni’s multiple comparisons ( g , h ). P values were adjusted for multiple comparisons. Source data are provided as a Source Data file. Data are representative of one ( i , j ), two ( d – h ), or three ( a , b ) independent experiments with similar results.

Article Snippet: LNP- HLA-EPs (0.5 μg and 10 μg per dose) or LNP -NC (10 μg) immunized HLA-A*02:01/DR1 transgenic mice or HLA-A*11:01/DR1 transgenic mice were injected intraperitoneally with an anti-mouse anti-CD8-α (IgG2b) mAb (Bio X Cell, Cat. No# BE0061, 100 μg/mouse, n = 5) or IgG2b isotype control mAb (Bio X Cell, Cat. No# BE0090, 100 μg/mouse, n = 5) on Days −1 and 0 of the challenge.

Techniques: Construct, Expressing, Flow Cytometry, Enzyme-linked Immunospot, Transgenic Assay, Infection, Variant Assay, Plaque Assay, Staining, Derivative Assay, Comparison

Journal: Nature Communications

Article Title: An mRNA-based T-cell-inducing antigen strengthens COVID-19 vaccine against SARS-CoV-2 variants

doi: 10.1038/s41467-023-38751-8

Figure Lengend Snippet: a Schematic diagram of the immunization, sample collection, and challenge schedule. b – d The humoral immune activation and antibody production after immunization. The lymph nodes collected ( n = 5) were analyzed for GC B cells and Tfh cells ( b ). SARS-CoV-2 RBD-specific IgG antibodies in the sera collected 21 days after the booster vaccination were detected with ELISA ( c ). The neutralizing antibodies against SARS-CoV-2 variants were detected with live viruses-based neutralization assays ( d ). The dashed lines represent that the LOD was 100 for ELISA, and that the lower limit of quantification was 8 for live virus neutralization. e – h The cellular immune response. Splenocytes of immunized HLA-A*02:01/DR1 transgenic mice ( n = 5) collected were restimulated ex vivo and subjected to IFN-γ ELISpot ( e ) (LOD = 5) and IL-4 ELISpot ( f ) (LOD = 2). The production of IFN-γ and TNF-α by CD8 + T cells ( g ) and CD4 + T cells was analyzed by an ICS ( h ). i , j The protective efficacy of the vaccine against the SARS-CoV-2 B.1.351 (Beta) and B.1.1.529 (Omicron BA.1) variants. Vaccinated HLA-A*02:01/DR1 transgenic mice ( n = 5) were challenged with the SARS-CoV-2 B.1.351 variant or B.1.1.529 variant, and viral titers at 4 days post-inoculation in the lung ( i -upper panel) and trachea ( i -bottom panel) tissues were determined by a plaque assay, with LODs being 20 and 100. The histopathological changes of the lungs at 4 days post-inoculation by the B.1.351 ( j -upper panel) or B.1.1.529 ( j -bottom panel) variant were evaluated after H&E staining. Images derived from one representative animal in each group with sites of inflammatory cell infiltration (black arrows), blood clots (blue arrows), and alveolar deformation (red arrows) are presented. Scale bar is 50 μm. Data ( b – j ) are shown as the mean ± S.E.M from individual mice. Source data are provided as a Source Data file. One ( b , j ) or two ( c – i ) independent experiments were performed with 2 technical replicates. Adjusted p values for statistical analysis were calculated via one-way ANOVA with Tukey’s multiple comparison post-hoc two-sided tests ( b , e – j ) or two-way ANOVA with Bonferroni’s multiple comparisons ( c , d ).

Article Snippet: LNP- HLA-EPs (0.5 μg and 10 μg per dose) or LNP -NC (10 μg) immunized HLA-A*02:01/DR1 transgenic mice or HLA-A*11:01/DR1 transgenic mice were injected intraperitoneally with an anti-mouse anti-CD8-α (IgG2b) mAb (Bio X Cell, Cat. No# BE0061, 100 μg/mouse, n = 5) or IgG2b isotype control mAb (Bio X Cell, Cat. No# BE0090, 100 μg/mouse, n = 5) on Days −1 and 0 of the challenge.

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Neutralization, Virus, Transgenic Assay, Ex Vivo, Enzyme-linked Immunospot, Variant Assay, Plaque Assay, Staining, Derivative Assay, Comparison

Journal: Nature Communications

Article Title: An mRNA-based T-cell-inducing antigen strengthens COVID-19 vaccine against SARS-CoV-2 variants

doi: 10.1038/s41467-023-38751-8

Figure Lengend Snippet: a Schematic diagram of the immunization, sample collection, and challenge schedule. b – d The humoral immune response against SARS-CoV-2. SARS-CoV-2 RBD-specific IgG antibodies in the sera of macaques were detected by ELISA using the RBD of the SARS-CoV-2 B.1.351 variant ( b ). The mean ± S.E.M. of RBD-specific IgG titers from 5 biological replicates were plotted against the days post-initial immunization when the sera were collected. The dashed line represents the LOD. The neutralizing antibodies were detected with pseudovirus-based ( c ) and live virus-based ( d ) neutralization assays. Pseudoviruses of six representative variants, Wuhan-Hu-1 (wild-type), B.1.1.7 (Alpha), B.1.351 (Beta), P.1 (Gamma), B.1.617.2 (Delta) and B.1.1.529 (Omicron BA.1), and infectious viruses of the six variants were exploited in the assays. The dashed lines represent the lower limit of quantification of 50 and 8, respectively. e , f The Th1-biased cellular immune response. PBMCs collected 14 days post-booster immunization were re-stimulated ex vivo and subjected to IFN-γ ELISpot (LOD = 5) ( e ) and IL-4 ELISpot (LOD = 2) ( f ), and representative dots are shown (right panels). g , h The protective efficacy of the vaccine against SARS-CoV-2. Macaques ( n = 3) were challenged with 7.0 × 10 5 PFU of SARS-CoV-2 (GDPCC-nCoV84, Beta variant) 3 weeks post-booster immunization. The copy number of SARS-CoV-2 sgRNA was determined by qRT-PCR in throat swabs (left panel) and anal swabs (right panel) obtained on Days 0, 1, 3, 5, and 7 post-inoculation ( g ). The LOD was established based on the standard curves at 100 copies per swab. The subgenomic copies in selected lung lobes collected from all macaques 7 days post-inoculation were determined by qRT-PCR ( h ). Data ( c – h ) are shown as the mean ± S.E.M from individual macaques. Source data are provided as a Source Data file. Experiments were repeated twice independently with similar results. Both independent experiments contain 3 technical repeats ( b – h ). Statistical significance was calculated via one-way ANOVA with Tukey’s multiple comparison post-hoc two-sided tests ( e , f , h ) or two-way ANOVA with Bonferroni’s multiple comparisons ( b – d , g ). P values were adjusted for multiple comparisons.

Article Snippet: LNP- HLA-EPs (0.5 μg and 10 μg per dose) or LNP -NC (10 μg) immunized HLA-A*02:01/DR1 transgenic mice or HLA-A*11:01/DR1 transgenic mice were injected intraperitoneally with an anti-mouse anti-CD8-α (IgG2b) mAb (Bio X Cell, Cat. No# BE0061, 100 μg/mouse, n = 5) or IgG2b isotype control mAb (Bio X Cell, Cat. No# BE0090, 100 μg/mouse, n = 5) on Days −1 and 0 of the challenge.

Techniques: Enzyme-linked Immunosorbent Assay, Variant Assay, Virus, Neutralization, Ex Vivo, Enzyme-linked Immunospot, Quantitative RT-PCR, Comparison

Journal: Frontiers in Immunology

Article Title: Enhanced Therapeutic Efficacy of Combining Losartan and Chemo-Immunotherapy for Triple Negative Breast Cancer

doi: 10.3389/fimmu.2022.938439

Figure Lengend Snippet: Losartan combined with chemo-immunotherapy promotes immune-stimulation in orthotopic 4T1 breast tumors. (A) Representative fluorescence images of 4T1 tumor slices immunostained for (effector T cell marker CD8, green), CD86 (M1-like TAM marker, green), CD206 (M2-like TAM marker, red), CRT (ICD effect marker, red) and DAPI (cell nuclei, blue). (B–D) Representative flow cytometry plots showing the tumor immune cells, including CD8+ T cells (CD45+, CD3+, CD8+), M1-like TAMs (CD11b+, F4/80+, CD86+) and M2-like TAMs (CD11b+, F4/80+, CD206+) in tumors after different treatments. (E) Heat map plot of ELISA results. (F) Quantification of the level of CD8+ T cells by flow cytometry analysis (n = 5). (G) Ratio of M2-like TAM to M1-like TAM by flow cytometry analysis (n = 5). (H) Quantification of CRT positive staining signals from the images shown in (A) (n = 9). Cytokine quantification of the secretion of TNF-α (I) , IFN-γ (J) , IL-6 (K) in sera from mice after various treatments as indicated in the figure (n = 3). Scale bar = 100μm. Data are expressed as the mean ± SEM. Statistical significances were calculated via one-way ANOVA, * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001. ns, no significance.

Article Snippet: Staining for CD8 was done by incubation with the anti-CD8-α mouse mAb (dilution 1:200, Santa Cruz, USA) and Alexa Fluor ® 488-conjugated goat anti-mouse secondary antibody (dilution 1:500, Absin, China).

Techniques: Fluorescence, Marker, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Staining

Journal: Molecular Imaging

Article Title: 89 Zr Immuno-PET Imaging of Tumor PD-1 Reveals That PMA Upregulates Lymphoma PD-1 through NF κ B and JNK Signaling

doi: 10.1155/2022/5916692

Figure Lengend Snippet: Immunohistochemistry of EL4 tumor tissue of mice with and without PMA treatment. CD8 α staining showing tumor-infiltrating effector lymphocytes (a) and PD-1 staining of tumor cells and lymphocytes (b) in tumors of control and PMA-treated mice. Magnification, ×12.5 (left) and ×200 (right).

Article Snippet: Microsection slides of paraffin tumor tissue underwent overnight incubation at 4°C with primary anti-mouse CD8 α (CST, #98941S; 1 : 100) or PD-1 antibodies (Abcam, #ab214421; 1 : 500).

Techniques: Immunohistochemistry, Staining, Control

Journal: Immunity

Article Title: Targeted complement inhibition at synapses prevents microglial synaptic engulfment and synapse loss in demyelinating disease

doi: 10.1016/j.immuni.2019.12.004

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse mAb α-CD8 (clone 8/114B) , Thermo Fisher Scientific , Cat. #MA5–13473.

Techniques: Virus, Recombinant, Adjuvant, Electron Microscopy, Cloning, Plasmid Preparation, RNAscope, Multiplex Assay, In Situ, Sequencing, Generated, Software, Imaging