Cd11a clone m17 4

Manufactured by BioLegend

CD11a (clone M17/4) is a monoclonal antibody that binds to the αL subunit of the lymphocyte function-associated antigen-1 (LFA-1) complex. LFA-1 is a cell surface receptor involved in cell-cell adhesion and leukocyte trafficking.

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3 protocols using cd11a clone m17 4

In vivo T cell Immunophenotyping

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In vivo labeling of T cells with fluorescent antibodies was performed as previously described (Turner et al., 2014). Briefly, mice were administered, PE- or Alexa 647-conjugated anti-CD90.2 (anti-Thy1.2; clone 53-2.1, BioLegend), or PE-conjugated anti-CD45.2 antibody (clone 104, Biolegend), intravenously, and 7–10 minutes later lungs were perfused, dissected and digested in RPMI1640 medium with collagenase D, DNAse and Trypsin inhibitor for 1 hour at 37°C. Mediastinal lymph nodes and spleen were dissected and manually disrupted to generate single cell suspensions. Cell suspensions were stained with fluorescent-conjugated antibodies for CD4 (clones RM4-5, BD Biosciences, and GK1.5, eBioscience), CD8 (clone 53-6.7, BioLegend), CD11a (clone M17/4, BioLegend),CD11b (M1/70 eBioscience), CD11c (N418, eBioscience), CD25 (PC61.5, eBioscience), CD45 (30-F11, BioLegend), CD69 (clone H1.2F3, eBioscience), CD86 (GL-1, Biolegend), CD103 (clone 2E7, eBioscience) and Anti-Mouse MHC Class II (I-A/I-E) (clone M5/114.15.2, eBioscience) was performed according to manufacturers’ protocol. Stained cells were analyzed using the BD LSRII flow cytometer and flow-jo software (Treestar, Ashland, OR). Absolute cell numbers were determined by flow cytometry using CountBright Absolute Counting Beads (Invitrogen) according to manufacturer’s protocol.

Turner D.L., Goldklang M., Cvetkovski F., Paik D., Trischler J., Barahona J., Cao M., Dave R., Tanna N., D’Armiento J.M, & Farber D.L. (2018). Biased generation and in situ activation of lung tissue-resident memory CD4 T cells in the pathogenesis of allergic asthma. Journal of immunology (Baltimore, Md. : 1950), 200(5), 1561-1569.

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Multiparametric Flow Cytometry of Immune Cells

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At indicated time points post-infection, the spleen and mesenteric lymph nodes from male and female mice were collected and mechanically disrupted to obtain a single-cell suspension. RBC lysis buffer from BioLegend (catalog # 420301) was used to remove erythrocytes. Afterward, the cells were washed and incubated with TruStain fcX (CD16/CD32, Clone 93, BioLegend, catalog # 101320) to prevent non-specific binding, and the immune cells of interest were then stained with appropriate surface antibodies. The samples were subjected to analysis using a BD LSRFortessa flow cytometer in combination with FlowJo software (BD Biosciences). The following mouse antibodies, in various fluorochrome combinations, were utilized for the staining: CD4 (clone GK1.5, BioLegend, catalog #100412, #100406), CD8a (clone 53-6.7, BioLegend, catalog, #100707 #100711), CD11a (clone M17/4, BioLegend, catalog # 101106), CD62L (clone MEL-14, BioLegend, catalog #104438), CD49d (clone R1-2, BioLegend, catalog #103618).

Pattnaik A., Dhalech A.H., Condotta S.A., Corn C., Richer M.J., Snell L.M, & Robinson C.M. (2024). A viral-specific CD4+ T cell response protects female mice from Coxsackievirus B3 infection. Frontiers in Immunology, 14, 1327384.

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Comprehensive Immunophenotyping by Flow Cytometry

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The following antibodies were used in an appropriate combination of fluorochromes: B220 (clone RA3–6B2, BioLegend), Bcl-6 (clone K112–91, BD Biosciences), CD4 (clone GK1.5, BioLegend), CD8α (clone 53–6.7, BioLegend), CD11a (clone M17/4, BioLegend), CD19 (clone 1D3/CD19, BioLegend), CD21 (Clone 7E9, BioLegend), CD23 (Clone B3B4, BioLegend), CD49d (clone R1–2, BioLegend), CD86 (Clone GL-1, BioLegend), CXCR5 (clone L138D7, BioLegend), FoxP3 (clone FJK-16s, eBioscience), GL7 (clone GL7, BioLegend), IFN-γ (clone XMG1.2, BioLegend), IgD (clone 11–26c.2a, BioLegend), IL-2 (clone JES6–5H4, BioLegend), IL-5 (clone TRFK5 BioLegend), IL-17A (clone TC11–18H10.1, BioLegend), PD-1 (clone 29F.1A12, BioLegend), T-bet (clone 4B10, BioLegend), TNF-α (clone MP6-XT22, BioLegend), and appropriate isotype controls.

Pardy R.D., Gentile M.E., Carter A.M., Condotta S.A., King I.L, & Richer M.J. (2022). An Epidemic Zika Virus Isolate Drives Enhanced T Follicular Helper Cell and B Cell-Mediated Immunity. Journal of immunology (Baltimore, Md. : 1950), 208(7), 1719-1728.

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