Characterization of anti-HLA class II monoclonal antibody LGII-612.14 reacting with formalin fixed tissues

Temponi, Massimo; Kekish, Ulana; Hamby, Carl V.; Nielsen, Hella; Marboe, Charles C.; Ferrone, S

doi:10.1016/0022-1759(93)90300-v

Cited by 58 publications

(34 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The monoclonal antibody HC-10, which recognizes a determinant expressed on virtually all h 2 m-free HLA-B heavy chains and on h 2 m-free HLA-A10, HLA-A28, HLA-A29, HLA-A30, HLA-A31, HLA-A32, and HLA-A33 heavy chains, was developed and characterized as described (23,24). The monoclonal antibody LGII-612.14, which recognizes a monomorphic determinant expressed on the h chain of HLA-DR, and on -DQ and -DP antigens, was developed and characterized as described (25). The TAP1 (NOB1) and TAP2 (NOB2) monoclonal antibodies were developed and characterized as previously described (26).…”

Section: Methodsmentioning

confidence: 99%

Modulation of Signal Transducers and Activators of Transcription 1 and 3 Signaling in Melanoma by High-Dose IFNα2b

Wang

Edington

Rao

et al. 2007

Clinical Cancer Research

108

View full text Add to dashboard Cite

Purpose: The Janus-activated kinase/signal transducers and activators of transcription (STAT) pathway of IFN signaling is important to immunoregulation and tumor progression. STAT1plays a prominent role in the effector immune response, whereas STAT3 is implicated in tumor progression and down-regulation of the response to type IIFNs.The goal of this study was to understand the effects of high-dose IFNa2b (HDI) in relation to the balance of pSTAT1and pSTAT3. Experimental Design: We evaluated STAT1 and STAT3 jointly as mediators of IFNa effects in the setting of a prospective neoadjuvant trial of HDI, in which tissue samples were obtained before and after 20 doses of HDI therapy. Double immunohistochemistry for pSTAT1and pSTAT3 was done on paired fixed (9 patients) or frozen (12 patients) biopsies. Results: HDI was found to up-regulate pSTAT1, whereas it down-regulates pSTAT3 and total STAT3 levels in both tumor cells and lymphocytes. Higher pSTAT1/pSTAT3 ratios in tumor cells pretreatment were associated with longer overall survival (P = 0.032). The pSTAT1/pSTAT3 ratios were augmented by HDI both in melanoma cells (P = 0.005) and in lymphocytes (P = 0.022). Of the immunologic mediators and markers tested,TAP2 was augmented by HDI (but not TAP1and MHC class I/II). Conclusion: IFNa2b significantly modulates the balance of STAT1/STAT3 in tumor cells and host lymphocytes, leading to up-regulation of TAP2 and augmented host antitumor response. The pSTAT1/pSTAT3 ratio in tumor cells at baseline may serve as a useful predictor of clinical outcome in cutaneous melanoma; the modulation of this ratio may serve as a predictor of therapeutic effect.High-dose IFNa2b (HDI) is the only therapy that has shown a reproducible ability to prolong both relapse-free and overall survival of patients with resected high-risk, deep, primary, or lymph node metastatic melanoma. This therapy has consistently shown a capacity to reduce the hazard of relapse and mortality by 22% to 33% in multiple U.S. intergroup studies done over the past 20 years (1 -3). However, in the past decade, no further progress in the adjuvant therapy of melanoma has occurred, and the gap in our knowledge of the IFNa2b mechanism has proven to be a major impediment to further progress. A better understanding of the mechanism(s) of HDI in the adjuvant therapy of melanoma would enable more effective application of this therapy, and more intelligent strategies building upon this modality.The antitumor effects of IFNa2b include direct inhibition of tumor cell growth and the modulation of host immune response to tumor. The Janus-activated kinase/signal transducers and activators of transcription (STAT) signal pathway is one of the major mechanisms of IFNa2b action. Both human type I IFNa receptor chains 1 and 2 are required for the type I IFN -dependent signaling pathways. The intracellular domains of these receptors are associated with Janus-activated kinases, which are activated upon IFNa2b binding to its receptors. Janus-activated kinases then phosphorylate ...

show abstract

Section: Methodsmentioning

confidence: 99%

Modulation of Signal Transducers and Activators of Transcription 1 and 3 Signaling in Melanoma by High-Dose IFNα2b

Wang

Edington

Rao

et al. 2007

Clinical Cancer Research

108

View full text Add to dashboard Cite

show abstract

“…The Delta(Y)-specific mAbs SY-4 and SY-5, the MB-1(X)-specific mAb SJJ-3, the LMP2-specific mAb SY-1, the LMP7-specific mAb HB-2, the LMP10-specific mAbs TO-6 and TO-7, the calnexin-specific mAb TO-5, the calreticulin-specific mAb TO-11, the ERp57-specific mAb TO-2, the tapasin-specific mAb TO-3, the mAb HC-10 that recognizes a determinant expressed on all h 2 M-free HLA-B and C heavy chains and on h 2 M-free HLA-A10, HLA-A28, HLA-A29, HLA-A30, HLA-A31, HLA-A32, and HLA-A33 heavy chains, the h 2 M-specific mAb L368, the HLA-DR, HLA-DQ, and HLA-DP-specific mAb LGII-612.14, and the MHC class I-related molecule B (MICB)-specific mAb SJJ-5 were developed and characterized as described (18,19,(24)(25)(26). The TAP1-specific mAb NOB-1 and the TAP2-specific mAb NOB-2 are secreted by hybridomas derived from the fusion of murine myeloma cells P3-X63-Ag8.653 with splenocytes from BALB/c mice immunized with partial length TAP1 recombinant protein (434-735) and a keyhole limpet hemocyanin-conjugated TAP1 peptide (717-735) and with partial length TAP2 recombinant protein (316-703), respectively, by the strategy described elsewhere (19).…”

Section: Methodsmentioning

confidence: 99%

Association of Antigen-Processing Machinery and HLA Antigen Phenotype of Melanoma Cells with Survival in American Joint Committee on Cancer Stage III and IV Melanoma Patients

Anichini¹,

Mortarini²,

Nonaka

et al. 2006

Cancer Research

View full text Add to dashboard Cite

Because changes in the expression level of antigen-processing machinery (APM) components and HLA class I and II antigens in melanoma cells are expected to affect their interactions with the immune system of the host, we assessed the clinical relevance of quantitative variations in the expression of these molecules in melanoma lesions. Short-term (<10 in vitro passages) melanoma cell lines isolated from 85 American Joint Committee on Cancer (AJCC) stage III and IV patients were stained with APM component and HLA class I antigen-specific and HLA class II antigen-specific monoclonal antibodies and analyzed by flow cytometry. The phenotype of all tumors was characterized by intertumor and intratumor heterogeneity in the expression of all the markers and by significant correlations in the level of expression of markers belonging to the HLA class I antigen-processing and presentation pathway. Hierarchical clustering of the mean fluorescence intensity data defined two main clusters of tumors. The corresponding groups of patients differed significantly in the overall survival but not in other relevant clinical variables, including AJCC stage and therapy received after surgery. Cox regression analysis showed that B 2 -microglobulin and HLA class II antigen expression were significantly associated with patients' survival. This evidence was corroborated by the immunohistochemical analysis for HLA class II antigen expression of melanoma lesions from an unrelated group of 52 AJCC stage III and IV patients. These results suggest that quantitative variations in APM component and HLA expression in melanoma lesions from AJCC stage III and IV patients may have an effect on the clinical course of the disease. (Cancer Res 2006; 66(12): 6405-11)

show abstract

“…72 Colocalization of DNA and HLA class II molecules was studied by immunostaining using, in addition, an anti-HLA class II IgG (LGII-612.14) monoclonal antibody. 73 The grids were then rinsed in TBS, incubated for 1 h at room temperature with goat anti-mouse IgG conjugated with 15 nm gold particles (EM GAM 15, British BioCell, Cardiff, UK), washed three times for 30 min in TBS, washed again in distilled water, and counterstained with uranyl acetate and lead citrate. All samples were examined with a Zeiss EM902 electron microscope (Oberkochen, Germany).…”

Section: Immunoelectron Microscopymentioning

confidence: 99%

Spontaneous transgenesis of human B lymphocytes

et al. 2003

View full text Add to dashboard Cite

DNA can cross the cell membrane by natural means, but the functional relevance of this phenomenon has not been fully elucidated. Here, we analyzed spontaneous transgenesis of human B cells using plasmid DNA coding for a functional immunoglobulin (Ig) heavy chain gene under the control of a B-cell-specific promoter. Using polymerase chain reaction (PCR), reverse transcriptase-PCR, and flow cytometry in combination, spontaneous transgenesis was documented in Burkitt's lymphoma cell lines, Epstein-Barr virus-transformed cell lines, and peripheral blood B lymphocytes of the mature naïve phenotype (IgM þ /IgD þ /CD27 À ). By immunoelectron microscopy, the internalized DNA was seen in the lysosomes/late endosomes and in the cytosol proximal to the nucleus. Importantly, spontaneously transgenic B cells processed and presented to major histocompatibility complex (MHC)-restricted T lymphocytes a peptide expressed in the transgenic product. This is the first demonstration that primary B lymphocytes possess a program for the spontaneous internalization of DNA, which in turn imparts the cell with new immunological functions. As spontaneous transgenesis is obtained using a nonviral vector, does not require prior cell activation, and is not associated with chromosomal integration, the findings reported here open new possibilities for genetic manipulations of mature naïve B lymphocytes for therapy and vaccination.

show abstract

Characterization of anti-HLA class II monoclonal antibody LGII-612.14 reacting with formalin fixed tissues

Cited by 58 publications

References 32 publications

Modulation of Signal Transducers and Activators of Transcription 1 and 3 Signaling in Melanoma by High-Dose IFNα2b

Modulation of Signal Transducers and Activators of Transcription 1 and 3 Signaling in Melanoma by High-Dose IFNα2b

Association of Antigen-Processing Machinery and HLA Antigen Phenotype of Melanoma Cells with Survival in American Joint Committee on Cancer Stage III and IV Melanoma Patients

Spontaneous transgenesis of human B lymphocytes

Contact Info

Product

Resources

About