Journal: Frontiers in Immunology

Article Title: Unveiling spatial complexity in solid tumor immune microenvironments through multiplexed imaging

doi: 10.3389/fimmu.2024.1383932

Figure Lengend Snippet: Immunophenotyping panel for multiplexed tissue imaging of cancer.

Article Snippet: CD3 , REA1151 , 50 , 130-120-267 , FITC (APC) , Miltenyi Biotec.

Techniques: Imaging

Journal: Journal of Translational Medicine

Article Title: Niraparib exhibits a synergistic anti-tumor effect with PD-L1 blockade by inducing an immune response in ovarian cancer

doi: 10.1186/s12967-021-03073-0

Figure Lengend Snippet: The information of antibodies used for western blot, immunohistochemistry, and flow cytometry

Article Snippet: CD3 and isotype , Human , 5 μl of antibody per test , Elabscience, E-AB-F1001E , Flow cytometry.

Techniques: Western Blot, Immunohistochemistry, Flow Cytometry

Journal: Advanced Science

Article Title: Bispecific Nanosystems Enable Multieffector Immune Cell Retargeting for Hematologic Malignancy Therapy

doi: 10.1002/advs.202509103

Figure Lengend Snippet: A) This diagram depicts the dual‐targeting strategy of biHSNPs in facilitating tumor cell elimination. Each biHSNP features two functional arms: one specifically binds to a tumor‐associated antigen on lymphoma cells (CD19/CD20), while the other engages and activates immune effector cells (CD3/CD16). By bridging tumor cells and immune cells, this approach enhances immune‐mediated tumor cell lysis, leveraging the immune system's inherent ability to identify and eliminate abnormal cells for precise and efficient cancer cell killing. B) Schematic illustration of the T cell activation and tumor cell death mechanism mediated by biHSNPs.

Article Snippet: FITC anti‐human CD19 antibody[CB19] (catalog No.: E‐AB‐F1004C, species reactivity: human), FITC anti‐human CD20 antibody[BCA/B20] (Catalog No.: E‐AB‐F1045C, species reactivity: human) and FITC anti‐human CD3 antibody[OKT‐3] (catalog No.: E‐AB‐F1001C, species reactivity: human) were obtained from Elabscience Biotechnology Co.,Ltd. (China, Wuhan).

Techniques: Functional Assay, Lysis, Activation Assay

Journal: Advanced Science

Article Title: Bispecific Nanosystems Enable Multieffector Immune Cell Retargeting for Hematologic Malignancy Therapy

doi: 10.1002/advs.202509103

Figure Lengend Snippet: Schematic illustration of the chemical synthesis and characterization of antibody‐conjugated hollow silica nanoparticles. A) Synthetic pathway of PEGylated hollow silica nanoparticles. B) Fourier transform‐infrared spectroscopy (FT‐IR) characterization of HSNP@PEG (2). C) Proton nuclear magnetic resonance ( 1 H NMR) characterization of HSNP@PEG. D) DLS and TEM images of HSNP@PEG. E) DLS and TEM images of HSNP@PEG@NH 2 (3). F) Fluorescence spectrum of HSNP@PEG@NH 2 @FITC (9.62 µ m ), confirming successful FITC conjugation. G) DLS and TEM images of HSNP@PEG@NH 2 @FITC (4). H–K) DLS and TEM results for monospecific nanoparticles (HSNP αCD3 , HSNP αCD16 , HSNP αCD19 , HSNP αCD20 ). L–O) DLS and TEM results for bispecific nanoparticles (HSNP αCD3 + αCD19 , HSNP αCD3 + αCD20 , HSNP αCD16 + αCD19 , HSNP αCD16 + αCD20 ). All scale bars in the TEM images represent 100 nm. HSNP αCD3 , HSNP αCD16 , HSNP αCD19 and HSNP αCD20 : Monospecific HSNPs conjugated with a single type of antibody (anti‐CD3, anti‐CD16, anti‐CD19, or anti‐CD20, respectively). HSNP αCD3 + αCD19 , HSNP αCD3 + αCD20 , HSNP αCD16 + αCD19 and HSNP αCD16 + αCD20 : Bispecific HSNPs conjugated with two different types of antibodies (e.g., anti‐CD3 and anti‐CD19). Antibody types are indicated using lowercase Greek letters (e.g., αCD3).

Article Snippet: FITC anti‐human CD19 antibody[CB19] (catalog No.: E‐AB‐F1004C, species reactivity: human), FITC anti‐human CD20 antibody[BCA/B20] (Catalog No.: E‐AB‐F1045C, species reactivity: human) and FITC anti‐human CD3 antibody[OKT‐3] (catalog No.: E‐AB‐F1001C, species reactivity: human) were obtained from Elabscience Biotechnology Co.,Ltd. (China, Wuhan).

Techniques: Fourier Transform Infrared Spectroscopy, Spectroscopy, Nuclear Magnetic Resonance, Fluorescence, Conjugation Assay

Journal: Advanced Science

Article Title: Bispecific Nanosystems Enable Multieffector Immune Cell Retargeting for Hematologic Malignancy Therapy

doi: 10.1002/advs.202509103

Figure Lengend Snippet: Binding specificity and cell–cell linkage induced by biHSNPs. Target cells were incubated with FITC‐labeled, antibody‐coated HSNPs at 4 °C for 30 min, followed by flow cytometry analysis to confirm binding specificity. A–C) Depict cell–cell linkage induced by biHSNPs: A) Schematic illustration of cell‐cell complex formation mediated by biHSNPs. B) Confocal microscopy images showing stable junctional complexes between immune effector cells and tumor cells facilitated by HSNP αCD3 + αCD19 and HSNP αCD3 + αCD20 . C) Flow cytometry analysis of cell–cell complexes between pre‐stained Jurkat (green, CellTracker Green CMFDA) and Raji cells (red, CellTracker Red CMTPX), demonstrating effective linkage by biHSNPs. D) Verification of biHSNPs binding to CD3 + cells using PBMCs as the CD3 + cell line. E) Verification of biHSNPs binding to CD19 + /CD20 + cells using Raji cells as the target. F) Verification of biHSNPs binding to CD16 + cells using PBMCs as the CD16 + cell line. G) Verification of biHSNPs binding to CD19 + /CD20 + cells using Raji cells as the target.

Article Snippet: FITC anti‐human CD19 antibody[CB19] (catalog No.: E‐AB‐F1004C, species reactivity: human), FITC anti‐human CD20 antibody[BCA/B20] (Catalog No.: E‐AB‐F1045C, species reactivity: human) and FITC anti‐human CD3 antibody[OKT‐3] (catalog No.: E‐AB‐F1001C, species reactivity: human) were obtained from Elabscience Biotechnology Co.,Ltd. (China, Wuhan).

Techniques: Binding Assay, Incubation, Labeling, Flow Cytometry, Confocal Microscopy, Staining

Journal: Advanced Science

Article Title: Bispecific Nanosystems Enable Multieffector Immune Cell Retargeting for Hematologic Malignancy Therapy

doi: 10.1002/advs.202509103

Figure Lengend Snippet: A) Cytotoxicity and cytokine release assays of biHSNPs in luciferase‐transfected Raji cells. B) In vitro cytotoxicity of HSNP@PEG, HSNP αCD3 , HSNP αCD19 , and bispecific HSNP αCD3 + αCD19 . C) In vitro cytotoxicity of HSNP@PEG, HSNP αCD3 , HSNP αCD20 , and bispecific HSNP αCD3 + αCD20 . D,E) Cytokine secretion levels (IFN‐γ, IL‐2, and TNF‐α) for HSNP αCD3 , HSNP αCD19 , and HSNP αCD3 + αCD19 ; and HSNP αCD20 and HSNP αCD3 + αCD20 . F) In vitro cytotoxicity of HSNP@PEG, HSNP αCD16 , HSNP αCD19 , and bispecific HSNP αCD16 + αCD19 . G) In vitro cytotoxicity of HSNP@PEG, HSNP αCD16 , HSNP αCD20 , and bispecific HSNP αCD16 + αCD20 . H,I) Cytokine secretion levels (IFN‐γ, IL‐2, and TNF‐α) for HSNP αCD16 , HSNP αCD19 , HSNP αCD16 + αCD19 , HSNP αCD20 , and HSNP αCD16 + αCD20 . J) Cytotoxicity of combined bispecific nanoparticle treatments: HSNP αCD3 + αCD19 with HSNP αCD16 + αCD19 , and HSNP αCD3 + αCD20 with HSNP αCD16 + αCD20 . K) In vitro cytotoxicity of HSNP αCD3 + αCD19 combined with HSNP αCD16 + αCD19 , and HSNP αCD3 + αCD20 combined with HSNP αCD16 + αCD20 . Data are presented as mean ± SD ( n ≥ 3), Statistical differences in B–I) were analyzed by One‐way ANOVA and K) were analyzed by Student's t‐test and the statistical significance is indicated as ** P ≤ 0.01 and *** P ≤ 0.001.

Article Snippet: FITC anti‐human CD19 antibody[CB19] (catalog No.: E‐AB‐F1004C, species reactivity: human), FITC anti‐human CD20 antibody[BCA/B20] (Catalog No.: E‐AB‐F1045C, species reactivity: human) and FITC anti‐human CD3 antibody[OKT‐3] (catalog No.: E‐AB‐F1001C, species reactivity: human) were obtained from Elabscience Biotechnology Co.,Ltd. (China, Wuhan).

Techniques: Luciferase, Transfection, In Vitro

Journal: Journal of Neuropathology & Experimental Neurology

Article Title: Lack of Major Histocompatibility Complex Class I Upregulation and Restrictive Infection by JC Virus Hamper Detection of Neurons by T Lymphocytes in the Central Nervous System

doi: 10.1097/nen.0000000000000218

Figure Lengend Snippet: FIGURE 1. T cells are present in white matter (WM), gray-white junction (GWJ), gray matter (GM), cerebellar granular layer (GL), and molecular layer (ML) areas harboring JCV-infected cells. Triple (A) and quadruple (BYD) immunostaining assays for CD8 (brown), CD3 (gray blue), T antigen (T Ag, purple), and VP1 (cyan-blue) show that numerous CD8-positive T cells (brown, filled circle arrow) are present in the vicinity of JC virus (JCV)Yinfected cells expressing T Ag (purple, arrowheads) in partially burnt out lesions located in the GWJ (A) square 1, magnified in inset 1 of an HIV-seropositive patient with progressive multifocal leukoencephalopathy. Rare presumed CD4-positive T cells (CD3 positive/CD8 negative, gray blue, asterisk in inset 1) are also present. The PML lesions extend partially in the GM (A, square 2, magnified in inset 2) where cells expressing T Ag can be seen (purple, arrowheads) as well as a few CD8-positive T cells located in the vessels (brown, empty circle arrows, inset 2). In B, the presence of CD8-positive and frequent presumed CD4-positive T cells in the GM of a JCV encephalopathy (JCVE) patient harboring numerous JCV-infected cells expressing T Ag and VP1. Numerous CD8-positive T cells in the parenchyma and blood vessels (brown, filled and empty circle arrows, respectively), presumed CD4-positive T cells (gray blue, asterisk), and JCV-infected cells expressing T Ag (purple, arrowhead) or VP1 (cyan-blue, arrow) are visible in the inset. An isolated cerebellar lesion at the border of the WM and GL of a JCV granule cell neuronopathy (JCV GCN) patient with chronic lymphocytic leukemia (C) and the magni- fication of the square in D show that CD8-positive T cells (brown, filled circle arrows) are present in the parenchyma in areas with JCV-infected cells expressing T Ag (purple, arrowheads) and/or VP1 (cyan-blue, arrows). Aggregates of CD8 on JCV-infected cells expressing VP1 are also evident. Scale bars = (AYC) 500 Km; (D) 50 Km.

Article Snippet: PAB597 m NA JCV VP1 SV40 T Ag (v-300); sc-20800 r IgG JCV T Ag Santa Cruz Biotechnology, Santa Cruz, CA MAP-2; HM-2; M4403 m IgG1 Neurons Sigma, St. Louis, MO MAP-2; ab5622 r IgG Neurons Chemicon, Temecula, CA MAP-2; LS-B290 ch IgY Neurons Lifespan Bioscience, Seattle, WA MAP-2; ab5392 ch IgY Neurons Abcam, Cambridge, MA NeuN (FOX3); ab104225 r IgG Neurons Abcam NeuN (FOX3); ABN90P gp IgG Neurons EMD Millipore, Billerica, MA GFAP; M0761; 6 F2 m IgG1 Astrocytes Dako, Carpinteria, CA GFAP; Z0334 r IgG Astrocytes Dako GFAP; ab4674 ch IgY Astrocytes Abcam GFAP; ab90601 sh IgG Astrocytes Abcam GFAP; 173004 gp IgG Astrocytes Synaptic Systems, Göttingen, DE CNPase; C5922; 11-5B m IgG1 Myelin/Oligo Sigma CNPase; ab18527 r IgG Myelin/Oligo Abcam CNPase; ab50739 ch IgY Myelin/Oligo Abcam CNP; HPA023266 r IgG Myelin/Oligo Sigma CD8; 1A5; VP-C325 m IgG1 CD8 Vector Laboratories, Burlingame, CA CD8 (144B); ab17147 m IgG1 CD8 Abcam CD3; CD3-12; OASA11552 rat IgG CD3 Aviva Systems Biology, San Diego, CA CD3; CD3-12; MCA1477 rat IgG CD3 AbD Serotec, Raleigh, NC CD68; M0814; KP1 m IgG1 Monocytes/ Dako Macrophages Granzyme B (11 F1); VP-G812 m IgG2a Granzyme B Vector Laboratories Perforin (5B10); VP-P967 m IgG1 Perforin Vector Laboratories MHC I (F-3), sc-55582 m IgG2a MHC I Santa Cruz Biotechnology MHC I (H-300), sc-25619 r IgG MHC I Santa Cruz Biotechnology HLA-A (EP1395Y); ab52922 r IgG MHC I Abcam ch, chicken; CNP and CNPase, CNP, 2¶,3¶-cyclic-nucleotide 3¶-phosphodiesterase; GFAP, glial fibrillary acidic protein; JCV, JC virus; m, mouse; MAG, myelin-associated glycoprotein; MAP-2, microtubule-associated protein 2; MOG, myelin-oligodendrocyte glycoprotein; oligo, oligodendrocytes; r, rabbit; SV40 T Ag, Simian virus 40 T antigen.

Techniques: Infection, Immunostaining, Virus, Expressing, Isolation

Journal: Journal of Neuropathology & Experimental Neurology

Article Title: Lack of Major Histocompatibility Complex Class I Upregulation and Restrictive Infection by JC Virus Hamper Detection of Neurons by T Lymphocytes in the Central Nervous System

doi: 10.1097/nen.0000000000000218

Figure Lengend Snippet: FIGURE 3. CD8-positive T cells aggregate mostly around cells with intact nuclei, expressing JC virus (JCV) T antigen (T Ag) and VP1. (A, B) Quadruple immunostaining for CD8 (brown), CD3 (gray blue), T Ag (purple), and VP1 (cyan-blue) done on an HIV- seronegative PML patient with chronic lymphocytic leukemia (A) and an HIV-seropositive PML patient (B). In PML lesions of the cerebral gray-white junction (GWJ) and gray matter (GM) (A, inset), where JCV-infected cells expressing VP1 are present (cyan-blue arrows), there are numerous parenchymal CD8-positive T cells (brown, black circle arrow). They are located mostly at the leading edge of infection where newly infected cells have still intact nuclei and express T Ag alone (purple, arrowhead) or T Ag and VP1 (cyan-blue arrows). In JCV-infected cerebellum (B), only rare CD8-positive T cells (brown, filled circle arrow) colocalize in the GL with hundreds of JCV-infected cells expressing T Ag (inset, purple, arrowheads), whereas more numerous CD8-positive T cells and some CD4-positive T cells (gray-blue, asterisk) colocalize in the contiguous WM with a few JCV-infected cells expressing VP1 (cyan-blue, arrow). (CYE) Triple immunofluorescence (IFA) staining for CD3, T Ag, and VP1 done in the cerebrum of an HIV-seronegative PML patient (C), an HIV-seropositive PML patient (D), and a JCV encephalopathy (JCVE) patient (E). The IFA immunostains confirm that T cells are absent in an area with T Ag only (C), whereas T cells are mostly present in areas where JCV-infected cells have intact nuclei with coexpression of T Ag and VP1 (pink, arrows in squares in D and E). Separate color channels are shown in insets. Scale bars = (A, B) 250 Km; (CYE) 100 Km.

Article Snippet: PAB597 m NA JCV VP1 SV40 T Ag (v-300); sc-20800 r IgG JCV T Ag Santa Cruz Biotechnology, Santa Cruz, CA MAP-2; HM-2; M4403 m IgG1 Neurons Sigma, St. Louis, MO MAP-2; ab5622 r IgG Neurons Chemicon, Temecula, CA MAP-2; LS-B290 ch IgY Neurons Lifespan Bioscience, Seattle, WA MAP-2; ab5392 ch IgY Neurons Abcam, Cambridge, MA NeuN (FOX3); ab104225 r IgG Neurons Abcam NeuN (FOX3); ABN90P gp IgG Neurons EMD Millipore, Billerica, MA GFAP; M0761; 6 F2 m IgG1 Astrocytes Dako, Carpinteria, CA GFAP; Z0334 r IgG Astrocytes Dako GFAP; ab4674 ch IgY Astrocytes Abcam GFAP; ab90601 sh IgG Astrocytes Abcam GFAP; 173004 gp IgG Astrocytes Synaptic Systems, Göttingen, DE CNPase; C5922; 11-5B m IgG1 Myelin/Oligo Sigma CNPase; ab18527 r IgG Myelin/Oligo Abcam CNPase; ab50739 ch IgY Myelin/Oligo Abcam CNP; HPA023266 r IgG Myelin/Oligo Sigma CD8; 1A5; VP-C325 m IgG1 CD8 Vector Laboratories, Burlingame, CA CD8 (144B); ab17147 m IgG1 CD8 Abcam CD3; CD3-12; OASA11552 rat IgG CD3 Aviva Systems Biology, San Diego, CA CD3; CD3-12; MCA1477 rat IgG CD3 AbD Serotec, Raleigh, NC CD68; M0814; KP1 m IgG1 Monocytes/ Dako Macrophages Granzyme B (11 F1); VP-G812 m IgG2a Granzyme B Vector Laboratories Perforin (5B10); VP-P967 m IgG1 Perforin Vector Laboratories MHC I (F-3), sc-55582 m IgG2a MHC I Santa Cruz Biotechnology MHC I (H-300), sc-25619 r IgG MHC I Santa Cruz Biotechnology HLA-A (EP1395Y); ab52922 r IgG MHC I Abcam ch, chicken; CNP and CNPase, CNP, 2¶,3¶-cyclic-nucleotide 3¶-phosphodiesterase; GFAP, glial fibrillary acidic protein; JCV, JC virus; m, mouse; MAG, myelin-associated glycoprotein; MAP-2, microtubule-associated protein 2; MOG, myelin-oligodendrocyte glycoprotein; oligo, oligodendrocytes; r, rabbit; SV40 T Ag, Simian virus 40 T antigen.

Techniques: Expressing, Virus, Immunostaining, Infection, Staining

Journal: Journal of Neuropathology & Experimental Neurology

Article Title: Lack of Major Histocompatibility Complex Class I Upregulation and Restrictive Infection by JC Virus Hamper Detection of Neurons by T Lymphocytes in the Central Nervous System

doi: 10.1097/nen.0000000000000218

Figure Lengend Snippet: FIGURE 5. CD8-positive T cells colocalize with JC virus (JCV)Yinfected glia rather than neurons in mixed areas. (A) Triple immu- nostaining for CD8 (brown), microtubule-associated protein 2 ([MAP-2] gray blue), and T antigen (T Ag) (purple) of the cerebrum of a HIV-seropositive patient with progressive multifocal leukoencephalopathy (PML).CD8-positive T cells (brown, black circle arrow) are common in the gray-white junction (GWJ) area (inset 1) in the vicinity of JCV-infected glia (MAP-2 negative) expressing T Ag (purple, arrowhead). Inset 2 shows that the isolated JCV-infected cells, which express T Ag alone (purple, arrowheads) and are located in the area free of CD8-positive T cells of the gray matter (GM), are in fact neurons (gray blue, arrowheads, MAP-2 positive)., (B) Triple immunofluorescence (IFA) staining for VP1 (inset 1), MAP-2 (inset 2), and granzyme B (inset 3) in the cerebrum of an HIV-seropositive PML patient shows that the CD8-positive T cells of the GWJ produce granzyme B when they are in contact with productively JCV-infected glia (MAP-2 negative, arrows). (C) Triple immunostaining for granzyme B (brown), MAP-2 (gray blue), and T Ag (purple) of the cerebrum of a JCV encephalopathy (JCVE) patient showing a T cell (black circle arrow) releasing its granzyme B (double arrow) in a JCV-infected neuron. (D) Quadruple IFA for CD3, CD8, granzyme B, and VP1 done on the same patient shown in C show numerous CD3-positive/CD8-positive T cells (yellow-red, filled circle arrows) producing granzyme B (white plus sign) as well as some presumed CD4-positive T cells (CD3 positive/CD8 negative, asterisk) devoid of granzyme B near VP1- expressing cells. A granzyme-producing CD3-positive/CD8-positive T cell (full circle arrow) apposed to a JCV-infected cell of neuronal phenotype expressing VP1 (arrow) is visible at the right of the inset. WM, white matter. Scale bars = (A) 1 mm; (BYD) 50 Km.

Article Snippet: PAB597 m NA JCV VP1 SV40 T Ag (v-300); sc-20800 r IgG JCV T Ag Santa Cruz Biotechnology, Santa Cruz, CA MAP-2; HM-2; M4403 m IgG1 Neurons Sigma, St. Louis, MO MAP-2; ab5622 r IgG Neurons Chemicon, Temecula, CA MAP-2; LS-B290 ch IgY Neurons Lifespan Bioscience, Seattle, WA MAP-2; ab5392 ch IgY Neurons Abcam, Cambridge, MA NeuN (FOX3); ab104225 r IgG Neurons Abcam NeuN (FOX3); ABN90P gp IgG Neurons EMD Millipore, Billerica, MA GFAP; M0761; 6 F2 m IgG1 Astrocytes Dako, Carpinteria, CA GFAP; Z0334 r IgG Astrocytes Dako GFAP; ab4674 ch IgY Astrocytes Abcam GFAP; ab90601 sh IgG Astrocytes Abcam GFAP; 173004 gp IgG Astrocytes Synaptic Systems, Göttingen, DE CNPase; C5922; 11-5B m IgG1 Myelin/Oligo Sigma CNPase; ab18527 r IgG Myelin/Oligo Abcam CNPase; ab50739 ch IgY Myelin/Oligo Abcam CNP; HPA023266 r IgG Myelin/Oligo Sigma CD8; 1A5; VP-C325 m IgG1 CD8 Vector Laboratories, Burlingame, CA CD8 (144B); ab17147 m IgG1 CD8 Abcam CD3; CD3-12; OASA11552 rat IgG CD3 Aviva Systems Biology, San Diego, CA CD3; CD3-12; MCA1477 rat IgG CD3 AbD Serotec, Raleigh, NC CD68; M0814; KP1 m IgG1 Monocytes/ Dako Macrophages Granzyme B (11 F1); VP-G812 m IgG2a Granzyme B Vector Laboratories Perforin (5B10); VP-P967 m IgG1 Perforin Vector Laboratories MHC I (F-3), sc-55582 m IgG2a MHC I Santa Cruz Biotechnology MHC I (H-300), sc-25619 r IgG MHC I Santa Cruz Biotechnology HLA-A (EP1395Y); ab52922 r IgG MHC I Abcam ch, chicken; CNP and CNPase, CNP, 2¶,3¶-cyclic-nucleotide 3¶-phosphodiesterase; GFAP, glial fibrillary acidic protein; JCV, JC virus; m, mouse; MAG, myelin-associated glycoprotein; MAP-2, microtubule-associated protein 2; MOG, myelin-oligodendrocyte glycoprotein; oligo, oligodendrocytes; r, rabbit; SV40 T Ag, Simian virus 40 T antigen.

Techniques: Virus, Infection, Expressing, Isolation, Staining, Triple Immunostaining

Journal: Journal of Neuropathology & Experimental Neurology

Article Title: Lack of Major Histocompatibility Complex Class I Upregulation and Restrictive Infection by JC Virus Hamper Detection of Neurons by T Lymphocytes in the Central Nervous System

doi: 10.1097/nen.0000000000000218

Figure Lengend Snippet: FIGURE 6. Major histocompatibility complex class I (MHC I) is upregulated in infiltrates and in parenchymal cells in JC virus (JCV)Yinfected areas. (AYF) Multiple immunofluorescence (IFA) staining images of CD3, MHC I, and VP1 (A, D, E), microtubule- associated protein 2 (MAP-2), MHC I, CD8, and VP1 (B, C), and CD68 and MHC I (F). A typical uninfected area in the temporal lobe of an HIV-seropositive patient with progressive multifocal leukoencephalopathy (PML) are shown in A and B. In those areas, cells expressing high level of MHC I are rare and restricted to vessels, parenchymal CD3-positive (A) and CD8-positive T cells (B) are absent and only rare CD8-positive T cells (B, empty circle arrow) are seen in blood vessels. Conversely, numerous cells expressing high levels of MHC I (red) are present in lesions of an HIV-seronegative patient with miliary PML (C) and a JCV encephalopathy (JCVE) patient (D) that harbor parenchymal CD8-positive (C) or CD3-positive (D) T cells (filled circle arrows) close to JCV-infected cells expressing VP1 (arrow and inset). Some of the cells expressing a high level of MHC I are infiltrates in the parenchyma and perivascular cuffs as shown by double positive for CD3-positive T cells and MHC I in a JCVE patient (white, E, inset) and for CD68-positive macrophages and MHC I in a pediatric HIV-seropositive PML patient (white, F, inset). Scale bar = 50 Km.

Article Snippet: PAB597 m NA JCV VP1 SV40 T Ag (v-300); sc-20800 r IgG JCV T Ag Santa Cruz Biotechnology, Santa Cruz, CA MAP-2; HM-2; M4403 m IgG1 Neurons Sigma, St. Louis, MO MAP-2; ab5622 r IgG Neurons Chemicon, Temecula, CA MAP-2; LS-B290 ch IgY Neurons Lifespan Bioscience, Seattle, WA MAP-2; ab5392 ch IgY Neurons Abcam, Cambridge, MA NeuN (FOX3); ab104225 r IgG Neurons Abcam NeuN (FOX3); ABN90P gp IgG Neurons EMD Millipore, Billerica, MA GFAP; M0761; 6 F2 m IgG1 Astrocytes Dako, Carpinteria, CA GFAP; Z0334 r IgG Astrocytes Dako GFAP; ab4674 ch IgY Astrocytes Abcam GFAP; ab90601 sh IgG Astrocytes Abcam GFAP; 173004 gp IgG Astrocytes Synaptic Systems, Göttingen, DE CNPase; C5922; 11-5B m IgG1 Myelin/Oligo Sigma CNPase; ab18527 r IgG Myelin/Oligo Abcam CNPase; ab50739 ch IgY Myelin/Oligo Abcam CNP; HPA023266 r IgG Myelin/Oligo Sigma CD8; 1A5; VP-C325 m IgG1 CD8 Vector Laboratories, Burlingame, CA CD8 (144B); ab17147 m IgG1 CD8 Abcam CD3; CD3-12; OASA11552 rat IgG CD3 Aviva Systems Biology, San Diego, CA CD3; CD3-12; MCA1477 rat IgG CD3 AbD Serotec, Raleigh, NC CD68; M0814; KP1 m IgG1 Monocytes/ Dako Macrophages Granzyme B (11 F1); VP-G812 m IgG2a Granzyme B Vector Laboratories Perforin (5B10); VP-P967 m IgG1 Perforin Vector Laboratories MHC I (F-3), sc-55582 m IgG2a MHC I Santa Cruz Biotechnology MHC I (H-300), sc-25619 r IgG MHC I Santa Cruz Biotechnology HLA-A (EP1395Y); ab52922 r IgG MHC I Abcam ch, chicken; CNP and CNPase, CNP, 2¶,3¶-cyclic-nucleotide 3¶-phosphodiesterase; GFAP, glial fibrillary acidic protein; JCV, JC virus; m, mouse; MAG, myelin-associated glycoprotein; MAP-2, microtubule-associated protein 2; MOG, myelin-oligodendrocyte glycoprotein; oligo, oligodendrocytes; r, rabbit; SV40 T Ag, Simian virus 40 T antigen.

Techniques: Immunopeptidomics, Virus, Staining, Expressing, Infection