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. 2020 Sep 16;8(2):e001052. doi: 10.1136/jitc-2020-001052

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© Author(s) (or their employer(s)) 2020. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.

This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See http://creativecommons.org/licenses/by-nc/4.0/.

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Clinical manifestation of binocular relapse of pro-B ALL after second CD19-CAR T-cell therapy. (A) Left eye before surgery. No intraocular structures are identifiable, the cornea appears clear but the anterior chamber is filled with a pus-like yellowish fluid, which resembles hypopyon in endophthalmitis. Note the hemorrhagic (reddish) appearance of the fluid which is not typically seen in infectious hypopyon. Additionally, the conjunctiva/sclera show a salmon-pink, ‘fleshy’ appearance, typically seen in conjunctival lymphoma. (B) Left eye after rinsing of the anterior chamber with extraction of the anterior chamber material. Now large dilated vessels on the surface of the iris can be identified at the periphery of the iris. The lens demonstrated a clear appearance. (C) CD3 and CD19 immunohistochemistry (IHC) of conjunctiva biopsy of the left eye. CD3⁺ T cells are concentrated at the surface of the conjunctiva. CD19⁺ cells show subepithelial localization and are widely observed within the connective tissue with additional perivascular accumulation. (D) CD3 and CD19 IHC from anterior ocular chamber lavage of the left eye. Most importantly, within anterior chamber lavage only CD19⁺ cells are present while in the CD3 IHC nucleated cells are CD3 negative. Black rectangles in left panel graphs illustrate the regions which are displayed in right panel graphs. for IHC, 4 µm sections of formalin-fixed paraffin-embedded tissue blocks were cut and antigen retrieval was carried out with microwave treatment and antigen target retrieval solution (Agilent technologies) for 64 min (pH 8.4). incubation for 28 min with either monoclonal rabbit anti-CD3 (1:150 dilution, clone Sp7, Zytomed) or monoclonal mouse anti-CD19 (1:20 dilution, clone EP169, Cellmarque) followed. Then, slides were incubated with a secondary anti-rabbit IgG antibody (MP-7401, ImmPress reagent kit, peroxidase-conjugated) followed by target detection using ultraView universal DAB detection kit (Ventana). BM, bone marrow; CAR, chimeric antigen receptor.