human cd4 isolation kit (Miltenyi Biotec)
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Miltenyi Biotec
human cd4 isolation kit
Human Cd4 Isolation Kit, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 97/100, based on 493 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/cd4+kit/CD4%2B+T+Cell+Isolation+Kit%2C+human/pmc13140695-37-0-6
Average 97 stars, based on 493 article reviews
Human Cd4 Isolation Kit, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 97/100, based on 493 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/cd4+kit/CD4%2B+T+Cell+Isolation+Kit%2C+human/pmc13140695-37-0-6
Average 97 stars, based on 493 article reviews
human cd4 isolation kit - by Bioz Stars,
2026-11
97/100 stars
Images
1) Product Images from "Role of Succinate Dehydrogenase in Age‐Related Th17 Inflammation"
Article Title: Role of Succinate Dehydrogenase in Age‐Related Th17 Inflammation
Journal: Aging Cell
doi: 10.1111/acel.70451
Figure Legend Snippet: Succinate dehydrogenase protein expression increases with age in CD4 + T cells. SDH expression in young (Y) and older (O) adults assessed via immunoblotting (a) and confocal microscopy (b). SDHA protein expression in CD4 + T cells treated with SDH inhibitor 3‐nitropropionic acid (3NP) and cell‐permeable diethyl succinate (DES); immunoblotting (c) and microscopy (d). SDHB protein expression after the different treatments (e). N = 3–4 (a–e). N = 3–4 indicates cells were obtained from either three or four individuals for each condition. Microscopy data are represented as cells in the field of view. At least 5–7 fields per slide were imaged at 63× magnification with oil immersion, on a Zeiss LSM 800 confocal microscope. In fields where numerous cells were observed, the mean fluorescence intensity of 3–4 cell groups was plotted as a single dot. Images were processed as described in the methods, and brightness was adjusted to improve clarity. Mann–Whitney Test or Kruskal–Wallis test with Dunn's post hoc test, * p < 0.05 vs. Y. # p < 0.05 vs. O or O + 3NP.
Techniques Used: Expressing, Western Blot, Confocal Microscopy, Microscopy, Fluorescence, MANN-WHITNEY
Figure Legend Snippet: Age‐induced dysregulation of TCA cycle metabolites fuel Th17 cytokine production. Waterfall plots showing ScRNA seq analysis of TCA cycle enzymes in CD4 + T cells from young (Y) and older (O) adults (a) ScRNA seq analysis of TCA cycle enzymes in Th17 subset of T cells (b). Cellular amounts of succinate (c) fumarate:succinate ratio (d) HIF1α protein in T cells from older adults (e) and HIF1α protein in T cells from younger adults after FH inhibition (f) N = 3, (a, b) N = 3–4, (c, d) N = 5–7, (e) N = 3, (f) adults in each group. One‐way ANOVA with Bonferroni test or Wilcoxon matched‐pair signed rank test. * p < 0.05 vs. O or Y.
Techniques Used: Inhibition
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![Continuous TGF-β exposure augmented CD25 expression and enabled superior IL-2–driven growth of long-term FOXP3 ± T cell cultures. (A, B) CD4 + T cells were purified from PBMCs and were activated for 3 d with PHA and 1 nM IL-2 in the presence or absence of 10 nM TGF-β to initiate two long-term lines. TGF-β–cultured T cells and control T cells were then passaged every 3 to 7 d in 1 nM IL-2 with or without 10 nM TGF-β respectively for a total of 38 d. Cells were stained with anti-CD3 PerCP-Cy5.5 (HIT3a), anti-CD4 BV421 (OKT4), anti-CD8 AF700 (SK1), anti-CD25 PE (M-A251), and anti-HLA-DR BUV395 (G46-6). Histograms (A) and bar charts (B) show flow cytometric analyses of cell size (forward scatter) together with CD3, CD4, CD25, and HLA-DR expression (n = 2). (C) Nonfractionated PBMCs (left) and purified CD4 + PBMC T cells (right) were activated and maintained as in panel A for a total of 20 d. To assess IL-2 responsiveness in proliferative assays, TGF-β–cultured T cells and control T cells were incubated with 10 pM–1 nM IL-2 (x-axis) with or without TGF-β in flat-bottomed 96-well plates. Cells were pulsed with [ 3 H]thymidine during the last 24 h of a 72-h culture (counts per minute [CPM]). (D) Independently derived T cell lines were initiated by a 3-d stimulation with PHA (rows 1, 2, 3, and 5) or without mitogen (autologous, row 4). T cells were cultured with 1 nM IL-2 in the presence or absence of 10 nM TGF-β (as designated) from day 0 until the day of analysis (day 12, 26, 38, or 47). Cells were stained with anti-FOXP3 clone PCH101 Alexa Fluor 488. (E) A nonfractionated T cell line was initiated as in panel A followed by continuous propagation in IL-2 and TGF-β for 19 d. Cells were gated on CD3 + and stained with the anti-FOXP3 clone 150D Alexa Fluor 488. Lines used in these experiments were CL2 (A, B), CL1 (C), NL1 (D, rows 1 and 2), CL1 (D, row 3), CL4 (D, row 4), CL2 (D, row 5), and P2 (E). * P < 0.05, ** P < 0.01. FMO, fluorescence minus one; FSC-A, forward scatter area; ns, not significant.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_6443/pmc13686443/pmc13686443__vkag274f1.jpg)
![A neutralizing anti-CD4 mAb inhibited autologous MHCII-restricted autoantigen recognition and IL-2–dependent growth of CD4 ± T-APCs. (A–G) Distinct CD4 + purified (A, D–G) and CD8 + purified (B) T-APC lines were initiated by a 3-d stimulation with PHA in the presence of 1 nM IL-2 and 10 nM TGF-β. Cells were passaged every 3 to 7 d in the presence of TGF-β and IL-2 for a total of 22 d (A), 80 d (B), 10 to 15 d (D-E), 78 d (F), 23 d (G, left two panels), or 36 d (G, right two panels). Human T cells (A, B, D–G) (50,000 cells/well, unless designated otherwise) or mouse SJL-PLP.1 T cells (C) (10,000 cells/well) were cultured with 10 pM, 100 pM, or 1 nM IL-2 (x-axis) in the presence or absence of anti-CD4 mAbs (OKT4 or RPA-T4) or isotype controls (MPC-11 or MOPC-21) in the presence or absence of 10 nM TGF-β. mAb concentrations were 20 μg/mL unless designated as 10 μg/mL. Cultures were pulsed with [ 3 H]thymidine during the last 24 h of a 5-d or 6-d culture unless designated otherwise. T-APC lines used in these experiments were CL5 (A, D), P2 (B), murine SJL-PLP.1 (C), CL6 (E, F), or CL7 (G). Two-way analysis of variance with a Tukey’s multiple comparisons test was used to assess significance. *** P < 0.001, and **** P < 0.0001 for comparisons of the designated mean value(s) at the same IL-2 concentration. CPM, counts per minute; ns, not significant.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_6443/pmc13686443/pmc13686443__vkag274f3.jpg)
![T-APC lines presented agonistic self-antigens and elicited polyclonal Tcon responses while seeding new T-APC lines. (A) JL3 Tcons were derived from freshly isolated PBMCs that were activated with Con-A and 1 nM IL-2 for 4 to 5 d. Tcons were passaged in 1 nM IL-2 in cRPMI media every 4 to 7 d for an additional 29 d. JL2.C5 T-APCs were derived in the presence of GM-CSF, IL-4, IL-2, and TGF-β (1 nM each). JL2.C5 cells were propagated in IL-2 and TGF-β for a total of 92 d (total expansion 10 7.4 -fold). Cells were stained with CD3 BV510 (SK7), CD4 BUV496 (SK3), CD8 Spark Blue 574 (SK1), CD25 PE-Fire 700 (MA251), HLA-DR Spark PLUS UV395 (L243), and CD40L PE (24-31). (B) JL2.C5 T-APCs (day 102) and JL3 Tcons (day 41 postactivation) were cultured for 24 h in the presence or absence of the anti-MHCII mAb L243 or the isotype control (10 μg/mL). (C) Similarly, JL2.C5 T-APCs (day 136) were cultured for 24 h in the presence or absence of anti-MHCII L243, anti-CD40L 5C8, or the isotype control (10 μg/mL). (D, E) Shown are the growth curves for the JL2.C5 T-APCs and the JL3 Tcon lines. (F) Designated densities (x-axis) of senescent nonexpanding JL2.C5 T-APCs (day 129) were added to 96-well plates with JL3 Tcons (day 16, 10 5 /well) in LymphoOne and 1 nM IL-2. Cells were passaged into 24-well plates on day 11. JL3 Tcon expansion was enumerated on day 18 with CountBright Absolute Counting Beads. (G) Cultures derived from panel F were expanded to T25 flasks, and after 2 to 4 wk 3 of these 18 cultures exhibited blastogenic T cell outgrowth. The initiation of the JL3 Tcon/JL2.C5 T-APC coculture (F) was designated as day 0 for initiation of the JL3.B1, JL3.C2, and JL3.T1 lines. (H) To assess whether the JL3.C2 T cells exhibited T-APC activity, JL3 Tcons (10 5 /well, day 39 out of activation) were cultured with designated densities (x-axis) of irradiated JL3.C2 T-APCs (5 Gy) for 5 d in the absence of exogenous mitogen or antigen. Cultures were pulsed with [ 3 H]thymidine during the last 48 h of culture. (I) T-APC lines CL2.2G4 and JL2.C6 and JL3 Tcon lines were compared with the nascent JL3.C2 T-APC line (day 32) for CD40L expression. (J) JL3.C2 T-APCs were cultured without exogenous TGF-β through day 58 and then were cultured with or without 1 nM TGF-β for 4 d. Cells were then subjected to a 10-day passage in duplicate cultures with 1 nM IL-2 in the presence or absence of 1 nM TGF-β without and without 10% heat-inactivated FBS, with manual counts on days 1, 3, 5, and 7 to 11. Cultures were supplemented with 1 nM IL-2 on day 5. (K) CD4 + JL3.C2 (day 70), CD8 + P3 T-APCs (day 14), murine BW5147α/β − cells, or SeAx cells were cultured in triplicate in a 2-d proliferative assay ([ 3 H]thymidine pulse during the last 24 h) with 1 nM IL-2 in the presence of designated titrations of heat-inactivated FBS. (B, C, I) Data were assessed with 1-way analysis of variance with Sidak’s or Dunnett’s multiple comparison test, respectively. (F, H, J, K) Data were assessed with a 2-way analysis of variance with Sidak’s multiple comparison test. ** P < 0.01, *** P < 0.001, **** P < 0.0001.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_6443/pmc13686443/pmc13686443__vkag274f5.jpg)
![T-APC coculture with syngeneic polyclonal Tcons caused an initial activation followed by fratricidal collapse. (A–D) The JL3.C2 T-APC line (day 43, 10 4 /well) and JL3 Tcons (day 39 of rest, 10 5 /well) were cultured alone or together in 96-well plates with 1 nM IL-2 (n = 3). On day 5, cells were passaged to a 24-well plate with 1 nM IL-2. On day 9, cells were subjected to flow cytometric analysis of CD3 BV785 (SK7), CD4 BV421 (OKT4), CD8 Alexa Fluor 700 (SK1), CD25 FITC (M-A251), CD28 BV605 (CD28.2), CD74 PE (LN2), CD86 BV650 (FUN-1), CD134 PE-Cy5 (ACT35), and HLA-DR Spark Plus UV395 (L243). (A) Shown are representative dotplots of the Tcon-derived CD4 + JL3.T1 and JL3.C2 T-APC lines, the combination culture (JL3.C2 T-APCs + JL3 Tcons), and Tcons alone. (B) Mean MFI values are shown for CD4, DN, and CD8 gated cells. Statistical analyses were restricted to comparison of “Both” (i.e. JL3.C2/Tcon cocultures) versus Tcons alone to assess coculture-induced changes in the Tcon phenotype. (C) Cell yields (day 9) were assessed with CountBright Absolute Counting Beads to calculate cell/bead ratios. (D) Manual counting was also used to compare cell yields (day 9) relative to input values for T-APCs (10 4 /well) and Tcons (10 5 /well), given that DN and CD8 + single-positive Tcons comprised approximately 40% of cells by day 9. (E) Total cell yields based on manual cell counting were assessed on days 16, 20, and 24. (F) JL3.C2 T-APCs were cultured without TGF-β for 58 d and then split into parallel cultures with and without 1 nM TGF-β for 3 d. Cells were then tested in a 2-d proliferative assay ([ 3 H]thymidine pulse during the last 24 h) in the presence of 1 nM IL-2 at 10 5 /well (200 µL in LymphoOne) in the presence or absence of 10 µg/mL functional grade anti-CD95 IgM mAb (EOS9.1). (G) CD4 + JL3.C2 T-APCs, CD8 + P3 T-APCs, SeAx T cells, and BW5147α/β − T cells tested in a proliferative assay as described in panel F at 50,000 cells/well with designated dilutions of the EOS9.1 mAb. (E) Data were assessed with an unpaired 2-tailed t test. (C–F) Data were assessed with 1-way analysis of variance with a Tukey’s multiple comparisons test. (B, G) Data were assessed with a 2-way analysis of variance with Sidak’s or Tukey’s multiple comparison test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. CPM, counts per minute. Flow cytometric analyses showed that T-APCs activated resting conventional responder T cells, which subsequently acquired a T-APC phenotype. The activated responder T cells then eliminated the original T-APCs and later exhibited mutual killing, ultimately causing collapse of the culture. Additional experiments showed that T-APCs were highly susceptible to FAS-mediated cell death following treatment with an agonistic cross-linking anti-FAS IgM antibody.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_6443/pmc13686443/pmc13686443__vkag274f10.jpg)
