Journal: Journal of immunological methods

Article Title: Cross-reactivity of T cell-specific antibodies in the bank vole (Myodes glareolus).

doi: 10.1016/j.jim.2023.113524

Figure Lengend Snippet: Fig. 2. A) Tree showing phylogenetic relationships among rodent species discussed in the article. Tree topology was based on a rodent phylogenetic tree published by Steppan and Schenk (2017), with root scaled to the median divergence time between Myodes and Mus reported on Timetree (see the main text). B) Pairwise amino acid sequence identity (upper-diagonal, in blue) and similarity (lower diagonal, in grey) matrices for CD4, CD8α and CD8β molecules. Mus musculus – mouse, Rattus norvegicus – rat, Peromyscus maniculatus - eastern deer mouse, Sigmodon hispidus - hispid cotton rat, Cricetulus griseus - Chinese hamster, Mesocricetus auratus - Syrian hamster, Microtus ochrogaster - prairie vole, Myodes glareolus – bank vole, and Homo sapiens, human. For brevity, only the generic name is provided on the figure. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: In addition, because anti-CD4 and anti-CD8α cotton rat-specific mAbs are commercially available, CD4 and CD8A hispid cotton rat (Sigmodon hispidus) sequences were obtained from commercially available constructs (Cotton Rat CD4 VersaClone cDNA, cat# RDC1063, R&D Systems; Cotton Rat CD8 alpha (AAL55392) VersaClone cDNA, cat# RDC0871, R&D Systems), which had been used A M. Migalska et al. Journal of Immunological Methods 520 (2023) 113524 in the construction of said antibodies.

Techniques: Sequencing

Journal: Journal of immunological methods

Article Title: Cross-reactivity of T cell-specific antibodies in the bank vole (Myodes glareolus).

doi: 10.1016/j.jim.2023.113524

Figure Lengend Snippet: Fig. 3. Expression of the CD4, CD8α and LCK genes in three sorted populations of cells: “CD4+” - CD3 + CD4+; “CD8+” - CD3 + CD4-; “neg” - CD3-CD4-. Relative normalized expression levels were measured with ΔΔCt method, with calibrator sample (“Calib”) prepared by mixing RNA from the three cell populations. TBP (TATA box binding protein) was used as a reference gene.

Article Snippet: In addition, because anti-CD4 and anti-CD8α cotton rat-specific mAbs are commercially available, CD4 and CD8A hispid cotton rat (Sigmodon hispidus) sequences were obtained from commercially available constructs (Cotton Rat CD4 VersaClone cDNA, cat# RDC1063, R&D Systems; Cotton Rat CD8 alpha (AAL55392) VersaClone cDNA, cat# RDC0871, R&D Systems), which had been used A M. Migalska et al. Journal of Immunological Methods 520 (2023) 113524 in the construction of said antibodies.

Techniques: Expressing, Binding Assay

Journal: PLoS Pathogens

Article Title: Synergistic Induction of Interferon α through TLR-3 and TLR-9 Agonists Identifies CD21 as Interferon α Receptor for the B Cell Response

doi: 10.1371/journal.ppat.1003233

Figure Lengend Snippet: (A) Spleen cells from cotton rats were stimulated with Concanavalin A for 72 hours and lymphoblasts purified by centrifugation on a ficoll-gradient. Subsequently, cells were stained with antisera specific for the human interferon receptor to test for crossreactive binding to cotton rat cells and analyzed by flow cytometry. Antibody specific for human interferon alpha receptor-1 (IFNAR-1) did not react with cotton rat cells (left) whereas antibody specific for human interferon alpha receptor-2 (IFNAR-2) did (right). IFNR specific antibody is shown as black line, controls as gray areas. B. IFNAR-2 antibody was added to cotton rat spleen cells. One hour later different concentrations of a combination of ODN2216 and poly I:C was added. 24 hours later supernatant was harvested and tested for the presence of type I interferon. The data is combined from two independent experiments. Significant differences were seen between cells treated with the combination of ODN2216 and poly I:C alone, or the combination of ODN2216 and poly I:C with antibody. IFNAR-1 antibody (not reactive with cotton rat cells) had no effect. C. Cotton rat spleen cells were stimulated with ODN 2216 or poly I:C, with or without the addition of recombinant cotton rat interferon alpha. (One unit equals one picogram of type I interferon.) 24 hours later supernatant was harvested and tested for the presence of type I interferon. The data is combined from two independent experiments, and significance was compared to cells without type I interferon treatment. Statistical analysis was done by ANOVA (* p<0.05, ** p<0.01, *** p<0.001).

Article Snippet: Neutralizing goat antibodies against cotton rat interferon alpha and IL-6 were purchased from R&D systems.

Techniques: Purification, Centrifugation, Staining, Binding Assay, Flow Cytometry, Recombinant

Journal: PLoS Pathogens

Article Title: Synergistic Induction of Interferon α through TLR-3 and TLR-9 Agonists Identifies CD21 as Interferon α Receptor for the B Cell Response

doi: 10.1371/journal.ppat.1003233

Figure Lengend Snippet: A) The number of MeV-specific B cells was measured from bone marrow cells of MeV-immune cotton rats. The addition of ODN 2216 and poly I:C individually and in combination increased B cell numbers whereas the addition of sera neutralizing cotton rat interferon alpha and IL-6 reduced this stimulation. Each bar graph represents the mean ± SD of triplicate wells. B) Cotton rats were immunized intranasally (upper panel) or subcutaneously (lower panel) with MeV, or MeV with ODN 2216 and/or pI:C in the presence or absence of human MeV-specific IgG (neutralization titer of 100) which had been injected intraperitoneally one day before immunization. Sera were collected at seven weeks post vaccination and the titer of neutralizing antibody was determined by neutralization assay. Each bar graph represents the average titer of four animals ± SD. The experiment is representative of three experiments. Statistical analysis was done by ANOVA (* p<0.05, ** p<0.01, *** p<0.001).

Article Snippet: Neutralizing goat antibodies against cotton rat interferon alpha and IL-6 were purchased from R&D systems.

Techniques: Neutralization, Injection

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Respiratory syncytial virus G protein and G protein CX3C motif adversely affect CX3CR1+ T cell responses.

doi: 10.4049/jimmunol.176.3.1600

Figure Lengend Snippet: FIGURE 7. G protein affects the number of IL-4- and CX3CL1-expressing cells. Mice were i.n. challenged with 5 105 PFU of WT or dG virus. At days 6 p.i. (A and C) and 12 p.i. (B and D), spleen cells were stimu- lated in vitro with purified G protein (G, 1000 nM), WT virus (RSV, 106 PFU), Con A (25 mg/ml), cell-free un- infected VCL (106 PFU equivalent), or medium alone, and the number of IL-4 (A and B)- or CX3CL1 (C and D)-expressing cells (ELISPOTs) was determined. The data are presented as the mean SD of the number of ELISPOTs/106 spleen cells from three experiments us- ing three mice/infection/experiment.

Article Snippet: Spleen cells were isolated from naive, WT, or dG virus-infected mice at days 6 or 12 p.i., and cultured in triplicate at 1 106 cells/well in multiscreen polyvinylidene difluoride nitrocellulose plates (Millipore) coated with 4 mg/ml anti-mouse IL-4 Ab (clone 11B11; BD Pharmingen) or 4 mg/ml anti-mouse CX3CL1 (clone MAB581; R&D Systems).

Techniques: Expressing, Virus, In Vitro, Infection