Journal: Frontiers in Cell and Developmental Biology

Article Title: Mesenchymal Stromal Cell-Conditioned Medium for Skin Diseases: A Systematic Review

doi: 10.3389/fcell.2021.654210

Figure Lengend Snippet: Studies regarding mesenchymal stromal cell-conditioned medium for treating wounds in animal models.

Article Snippet: , Human amnion , Amniocentesis , Flow cytometry (CD73, CD45, CD31, CD105, CD44, CD34, CD90, CD29, SSEA-3, SSEA-4, EP-CAM, HLA-DR). Chondrogenic, osteogenic, and adipogenic differentiation , MSCs of passage 2 at 90% confluence were used. CM was collected , Murine. Full-thickness excisional skin wounds, 10 mm, on the dorsum , 50 μl subcutaneous injected into the surrounding tissues of the wound bed at four sites - PBS (control) - MSC-CM - Murine LOXL2 (5 μg, Sino Biologicals, China) ( n = 4/group) , 12 , Macroscopic appearance (photography), histology (HE, TC) , Wound sizes were significantly reduced in the LOXL2 and MSC-CM groups compared with controls ( p < 0.05) , MSC-CM and LOXL2 enhanced wound healing. Epidermis of the MSC-CM and LOXL2 group mice resembled normal skin and the keratinocytes were well organized and tightly arranged. These treatments also significantly reduced fibrosis and improved keratinocyte proliferation. No adverse events were reported.

Techniques: Flow Cytometry, Expressing, Isolation, Injection, Transfection, Recombinant, Construct, Dissection, Transduction, Plasmid Preparation, Fluorescence, Marker, Cell Culture, Staining, Activation Assay, Migration, Enzyme-linked Immunosorbent Assay, Functional Assay, Generated, Immunofluorescence, Immunohistochemistry-IF, Immunoprecipitation

Journal: Oncotarget

Article Title: Tumor antigen CA125 suppresses antibody-dependent cellular cytotoxicity (ADCC) via direct antibody binding and suppressed Fc-γ receptor engagement

doi: 10.18632/oncotarget.19090

Figure Lengend Snippet: A . Biotinylated farletuzumab and its (Fab’) 2 fragment, but not its Fc fragment bind immobilized sCA125. 96-well plates were coated with 15 KU/mL sCA125 or human serum albumin (HSA) and probed with biotin-labeled farletuzumab, (Fab’) 2 or Fc fragments. B . Farletuzumab is able to compete for farletuzumab-biotin binding to sCA125. C . sCA125 suppresses Fc receptor binding to farletuzumab. Farletuzumab was incubated alone or with sCA125 or HSA and probed with biotinylated-CD16a, -CD32a or -CD64a Fc receptor and graphed as a percent inhibition of Fc receptor binding compared to control. sCA125 caused a significant decrease of CD16a binding to farletuzumab as compared to controls (46%, p < 0.0002). Reduction in farletuzumab binding to CD32a Fc receptors was also significant (12%, p < 0.003) while minimal inhibition was observed by sCA125 on farletuzumab binding to CD64a Fc receptor (1%, p = 0.772). Similar reactions were probed with anti-human IgG-HRP to confirm incubation of farletuzumab with sCA125 did not result in less farletuzumab binding to wells . D . BRA assay using CHO cells expressing human Fc activating receptors show only farletuzumab-CD16a interactions are disrupted by sCA125. E . Farletuzumab has a significant increase in binding to the mCA125 producing OVCAR parental as compared to mCA125-suppressed isogenic OVCAR-KD1 or mCA125-null CHO membranes. F . sCA125 can bind to subsets of other humanized and fully human mAbs. Seven purified mAbs were biotinylated and used to probe wells coated with 15 KU/mL sCA125 or HSA. While all antibodies were able to robuslty and equally bind their respective target antigen, only a subset were able to specifically bind sCA125 as compared to HSA controls. All ELISAs were done in at least triplicate and different lots of sCA125 or cell membrane preparations were used with similar results. In panels A and B *** p < 0.0001; **** p < 0.00007; ***** p < 0.000001, panels E and F *** p < 0.0001; **** p < 0.00001.

Article Snippet: Human CD16a-, CD32a- and CD64a-biotin probes were purchased from Sino Biological Inc. Biotinylated CA125 or HSA were generated as previously described [ ].

Techniques: Labeling, Binding Assay, Incubation, Inhibition, Expressing, Purification

Journal: The Journal of Allergy and Clinical Immunology

Article Title: ARGX-117, a therapeutic complement inhibiting antibody targeting C2

doi: 10.1016/j.jaci.2020.08.028

Figure Lengend Snippet: ARGX-117 does not interact with C1q (A) , CD16 (B) , or CD32a (C) , and binds stronger to FcRn than to wild-type (WT) IgG 1 at pH 6.0 (D). A, ELISA plates were coated with ARGX-117, incubated with serum, and C1q was detected with anti-C1q. B-D, Plates were coated with NeutrAvidin and incubated with biotinylated CD16 (B) , CD32a (C) , or FcRn (D) . Subsequently, plates were incubated with ARGX-117. Bound ARGX-117 was then detected with poly-HRP-labeled goat anti-human IgG antibody.

Article Snippet: Plates were then incubated for 1 hour with anti-human CD32a (His Tagged; Sino Biological) and incubated with serial dilutions of ARGX-117 or ARGX-18E2 for 1 hour.

Techniques: Enzyme-linked Immunosorbent Assay, Incubation, Labeling