sialic acid sa receptor expression  (Vector Laboratories)

Journal: Emerging Microbes & Infections

Article Title: Pandemic influenza A (H1N1) virus causes abortive infection of primary human T cells

doi: 10.1080/22221751.2022.2056523

Figure Lengend Snippet: H1N1 infected different subsets of primary human T cells heterogeneously. (A) Distribution of different subsets in infected human CD4 + T cells. Left, 0 h.p.i.; right, 16 h.p.i. Blue, naïve CD4 + T; green, follicular helper T (Tfh); wine, CD4 + cytotoxic T (CTL); rose pink, regulatory T (Treg). (B) Distribution of different subsets in infected human CD8 + T cells. Left, 0 h.p.i.; right, 16 h.p.i. Blue, naïve CD8 + T; green, stem cell memory T (T SCM ); wine, effector memory T (T EM ); rose pink, terminally differentiated effector T (T TE ); yellow, exhaust T (T EX ). (C) Infection rates among different primary human CD4 + and CD8 + T subsets. Blue, 0 h.p.i.; red, 16 h.p.i.; green dotted line, 5%. (D) mRNA expression levels of viral HA in naïve CD8 + T and CD8 + T EM from one healthy donor at eight time points. Results are represented as mean fold change ± SD and statistical significances were analysed using GraphPad Prism 8.0 through Student’s t -test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns = non-significant. (E) Immunofluorescent staining of naïve CD8 + T and CD8 + T EM which exposure to H1N1 respectively. Green, α-2,6-linked sialic acid receptors; pink, CD45RO; red, viral HA proteins; blue, DAPI (4′,6-diamidino-2-phenylindole).

Article Snippet: Primary antibodies to α-2,6-linked sialic acid receptors (Vector Laboratories®, FL-1301-2), CD45RO (abcam, [UCH-L1] ab23) and influenza H1N1 viral HA antibody (Sino Biological®, 11085-T54) was diluted 1:200 in blocking buffer (1% BSA) and incubated overnight at 4 °C.

Techniques: Infection, Expressing, Staining

Journal: Emerging Microbes & Infections

Article Title: Pandemic influenza A (H1N1) virus causes abortive infection of primary human T cells

doi: 10.1080/22221751.2022.2056523

Figure Lengend Snippet: Quantitative analysis of α-2,3- and α-2,6-linked sialic acid receptors in MDCK.2 and primary human T cells. (A) Flow cytometry analysis of broad-spectrum neuraminidases treated MDCK.2 cells or not with the lectin antibody concentrations of 5 and 20 μg/mL. Grey, unstained as controls; blue, α-2,6-linked sialic acid receptors; red, α-2,3-linked sialic acid receptors. (B) The comparison of α-2,6- and α-2,3-linked receptors in MDCK.2 cells from ATCC. (C) The comparison of α-2,6- and α-2,3-linked receptors of human CD14 + mononuclear/macrophages. (D) The comparison of α-2,6- and α-2,3-linked receptors of human total CD4 + T and total CD8 + T cells. (E) Purities of gating and MFI of receptors with the lectin antibody concentrations of 5 μg/mL in CD8 + T EM and naïve CD8 + T cells and the quantitative analysis of α-2,6- and α-2,3-linked sialic acid receptors between CD8 + T EM and naïve CD8 + T cells from five healthy donors. Both CD8 + T EM and naïve CD8 + T cells were stained with identical fluorescent antibodies. Red, naïve CD8 + T; blue, CD8 + T EM . Results of (B-E) are represented as mean fold change ± SD and statistical significances were analysed using GraphPad Prism 8.0 through two-tailed Student’s t -test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Primary antibodies to α-2,6-linked sialic acid receptors (Vector Laboratories®, FL-1301-2), CD45RO (abcam, [UCH-L1] ab23) and influenza H1N1 viral HA antibody (Sino Biological®, 11085-T54) was diluted 1:200 in blocking buffer (1% BSA) and incubated overnight at 4 °C.

Techniques: Flow Cytometry, Staining, Two Tailed Test