Intercellular Adhesion Molecule-1 Antigen Research Articles

ICAM1 antibody drug conjugates exert potent antitumor activity in papillary and anaplastic thyroid carcinoma

Treatment options for anaplastic thyroid cancer (ATC) and refractory papillary thyroid carcinoma (PTC) are limited and outcomes remain poor. In this study, we determined via bioinformatic expression analyses and immunohistochemistry staining that intercellular adhesion molecule-1(ICAM1) is an attractive target for ATC and PTC. We designed and engineered two ICAM1-directed antibody-drug conjugate (I1-MMAE and I1-DXd), both of which potently and selectively ablate multiple human ATC and PTC cell lines without affecting non-plastic cells invitro. Furthermore, I1-MMAE and I1-DXd mediated a potent tumor regression in ATC and PTC xenograft models. To develop a precision medicine, we also explored magnetic resonance imaging (MRI) as a non-invasive biomarker detection method to quantitatively map ICAM1 antigen expression in heterogeneous thyroid tumors. Taken together, this study provides a strong rationale for the further development of I1-MMAE and I1-DXd as promising therapeutic candidates to treat advanced PTC and ATC.

Treatment of adhesions after Achilles tendon injury using focused ultrasound with targeted bFGF plasmid-loaded cationic microbubbles

AbstractNonviral targeting technology has become promising as a form of gene therapy for diseases and injuries, such as Achilles tendon injuries. In this study, we used avidin–biotin bridge and positive–negative charge attractions to load the intercellular adhesion molecule-1 (ICAM-1) antibody and the basic fibroblast growth factor (bFGF) plasmid onto the surface of the microbubbles. The saturated loading capacity for 1.1 × 108microbubble was 6.55 ± 0.53 µg. We established the ICAM-1 antigen microenvironment using tumor necrosis factor-alpha-stimulated human umbilical vein endothelial cells and found the targeting ability of the prepared microbubblesin vitro.In vivo, we also found that the injected targeted bFGF gene microbubbles expressed the bFGF gene better when compared with that of the control group. Furthermore, we evaluated adhesions after Achilles tendon injuries in rabbits using hematoxylin and eosin and immunohistochemical (IHC) staining methods. The collagen fibers were properly arranged in the tendon, and there was greater cellularity inside the tendon sheath and a clearer boundary between the internal and external tendon sheath than that of the control group. IHC staining showed greater ICAM-1 expression inside the tendon sheath when compared with outside the tendon sheath. In conclusion, targeted microbubbles can be a useful carrier of genes to provide gene therapy for the prevention of adhesions after tendon injury.

Preparation and identification of anti-human ICAM-1 scFv

To screen the specific anti-human intercellular adhesion molecule-1 (ICAM-1) single chain fragment variable (scFv) using phage display library technology and to identify its biological activity. P1 peptide was used as antigen, and the phage antibodies against human ICAM-1 antigen were panned by four binding-eluting-amplifying cycles using Tomlinson I+J phage display library. After four rounds of selective enrichment screening, the positive clones were determined by PCR, enzyme linked immunosorbent assay (ELISA)-based antigenic cross reaction and Dot blotting. Then the binding specificity and biological activity of purified scFv were identified by Western blotting, competitive ELISA and cell adhesion inhibition assay respectively. Furthermore, four positive clones were first panned through P1 peptide coated-ELISA assay, and then J-A1 was obtained and identified by PCR, ELISA-based antigenic cross reaction and Dot blotting, which could show a specific binding between P1 peptide and human ICAM-1 protein antigen. Subsequently, the purified scFv showed a satisfactory specificity and anti-adhesive activity in competitive ELISA and the cell adhesion inhibition assay. The specific anti-human ICAM-1 scFv was prepared successfully from Tomlinson I+J phage display library, which pave the way for further application of anti-human ICAM-1 scFv for inflammation diseases therapeutics.

ICAM-1 depletion in the center of immunological synapses is important for calcium releasing in T-cells

T-cell activation requires the formation of the immunological synapse (IS) between a T-cell and an antigen-presenting cell (APC) to control the development of the adaptive immune response. However, calcium release, an initial signal of T-cell activation, has been found to occur before IS formation. The mechanism for triggering the calcium signaling and relationship between calcium release and IS formation remains unclear. Herein, using live-cell imaging, we found that intercellular adhesion molecule 1 (ICAM-1), an essential molecule for IS formation, accumulated and then was depleted at the center of the synapse before complete IS formation. During the process of ICAM-1 depletion, calcium was released. If ICAM-1 failed to be depleted from the center of the synapse, the sustained calcium signaling could not be induced. Moreover, depletion of ICAM-1 in ISs preferentially occurred with the contact of antigen-specific T-cells and dendritic cells (DCs). Blocking the binding of ICAM-1 and lymphocyte function-associated antigen 1 (LFA-1), ICAM-1 failed to deplete at the center of the synapse, and calcium release in T-cells decreased. In studying the mechanism of how the depletion of ICAM-1 could influence calcium release in T-cells, we found that the movement of ICAM-1 was associated with the localization of LFA-1 in the IS, which affected the localization of calcium microdomains, ORAI1 and mitochondria in IS. Therefore, the depletion of ICAM-1 in the center of the synapse is an important factor for an initial sustained calcium release in T-cells.

Abstract 2207: Using AFM to predict the specificity of tumor targeting nanomedicine in vivo

Abstract Most cancer target and biomarker studies focus on the identification of overexpressed genes or proteins of cancer cells. However, their potential feasibility as tumor-specific antigens targeted by antibody-guided nanomedicine has not consistently been based on quantitative assessments of tumor specificity and binding strength. Cancer cell membrane surfaces possess hundreds of different functionally versatile proteins (receptors and ligands) which are heterogeneously upregulated or downregulated to promote cancer development and progression. Selection of appropriate drug targets is pivotal to the successful development of tumor-targeted therapeutics because tumor-specific affinity of potential cancer targets is a key factor modulating cellular binding and uptake, and has the potential to translate to therapeutic bioactivity. To date, the quantitative measurements of cancer target antibody-antigen interactions are often limited by dynamic biophysical and biomechanical methods for probing such interactions. To address this challenge, we used flow cytometric analyses to compare 40 potential cancer cell surface target candidates for triple negative breast cancer (TNBC), an aggressive malignancy with limited treatment options. We selected intercellular adhesion molecule-1 (ICAM-1) as the “optimized” TNBC target for nanomedicine. Reasoning that adhesion forces between the ICAM-1 antibody and live human breast cancer cells could be used to guide nanomedicine delivery to TNBC tumors, we used atomic force microscopy (AFM) to evaluate the binding strength between the ICAM-1 antibody and antigen on live human TNBC cells and non-neoplastic cells. Adhesion forces between the ICAM-1 antibody and live human TNBC MDA-MB-231 cells (523 ± 113 pN) were significantly higher than those of non-neoplastic MCF10A cells (336 ± 33 pN). We next validated these findings by engineering ICAM-1-targeted immunoliposomes and then tested their TNBC-specific binding affinity and cytotoxicity on three TNBC cell lines. In vivo imaging experiments further confirmed that the ICAM-1-targeted immunoliposomes preferentially accumulated in orthotopic TNBC tumors, in comparison with nonspecific, IgG-conjugated liposomes. These data support the conclusion that ICAM-1 antibody-conjugated immunoliposomes containing chemotherapeutic drugs have the potential to preferentially target and destroy TNBC cells, and may also be applicable to a range of metastatic cancers that demonstrate an overexpression of ICAM-1. These findings also support the use of AFM as a valuable tool in the characterization of surface antigen-antibody interactions that predict in vitro and in vivo binding affinity. Acknowledgements: D. Auguste acknowledges the support of NIH 1DP2CA174495. M. Moses acknowledges the support of NIH R01CA185530 and the Breast Cancer Research Foundation. Citation Format: Peng Guo, Biran Wang, Daxing Liu, Jiang Yang, Marsha Moses, Debra Auguste. Using AFM to predict the specificity of tumor targeting nanomedicine in vivo. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2207.

Allograft rejection-related gene expression in the endothelial cells of renal transplantation recipients after cytomegalovirus infection

To explore the effects of cytomegalovirus (CMV) infection on rejection-related gene expression in the endothelial cells of renal transplantation recipients. Endothelial cells (ECs) were cultured and stimulated by a variety of factors: A, normal control group; B, inactivated human cytomegalovirus (HCMV) infection group; C, HCMV infection group; D, HCMV supernatant infection group; and E, ganciclovir HCMV group. Expression of intercellular adhesion molecule-1 (ICAM-1) and major histocompability complex (MHC) class I and class II antigens was detected by flow cytometry (FCM) and immunohistochemistry. We found characteristic CMV-infected ECs in this study. There were no significant differences among groups A, B and D (P>0.05). Although the expression levels of ICAM-1 were not significantly different between groups C and E (P>0.05), the ICAM-1 expression in these two groups was significantly higher than that in group A (P<0.05). ICAM-1 expression was detected in groups C and E, while there was no expression in groups A, B and D. Furthermore, there was no significant difference of ICAM-1 mRNA expression between groups C and E (P>0.05). Human leucocyte antigen (HLA)-ABC expression was detected in all the groups, while HLA-DR expression was only detected in groups C and E. There were no significant differences of HLA-ABC and HLA-DR expression among groups A, B and D (P>0.05). However, the HLA-ABC and HLA-DR expression levels in groups C and D were higher than those of the remaining groups previously reported (P<0.05). Meanwhile, the HLA-ABC and HLA-DR expression levels in group E were lower than those of group C (P<0.05). CMV could up-regulate the expression levels of ICAM-1 and MHC antigens, which was closely related to allograft rejection.

Detection of circulating intercellular adhesion molecule-1 antigen in malignant diseases

A new monoclonal antibody (MoAb) HA58 (IgG1) was prepared, which recognizes the binding site on the intercellular adhesion molecule-1 (ICAM-1) antigen to the lymphocyte function-associated antigen-1 (LFA-1). The double-determinant immunoassay (DDIA) was established with use of MoAb HA58 and another anti-ICAM-1, MoAb CL207, to detect the soluble, shedding ICAM-1 antigen. Human recombinant interferon-gamma (IFN-gamma) induced not only the expression of cell surface ICAM-1, but also the shedding ICAM-1 antigen in an IFN-gamma concentration-dependent and incubation-time-dependent manner. DDIA was applied to detect the shedding ICAM-1 antigen in the sera of patients with malignant or benign diseases. The incidence of positivity for ICAM-1 antigen in malignant diseases was higher than that in benign diseases or in healthy controls. Furthermore, the sera of cancer patients with liver metastasis showed higher levels of the shedding ICAM-1 antigen. These findings suggest that serum ICAM-1 antigen may be a useful marker to monitor tumor burden in cancer patients.

Circulating intercellular adhesion molecule-1 (ICAM-1) antigen in sera of patients with idiopathic pulmonary fibrosis.

Intercellular adhesion molecule-1 (ICAM-1), a member of immunoglobulin supergene family with a five-domain structure, is known to play an important role in inflammatory diseases. An ELISA was developed using two MoAbs against human ICAM-1 in order to detect the soluble shedding ICAM-1 antigen in sera. We measured levels of circulating ICAM-1 antigen in sera of patients with idiopathic pulmonary fibrosis (IPF), pulmonary sarcoidosis, hypersensitive pneumonitis, bacterial and mycoplasmal pneumonia, and inflammatory diseases of other organs. The results clearly demonstrated that IPF had significantly high levels of circulating ICAM-1 in sera as compared with other disorders or normal controls. Moreover, immunohistochemical analysis with MoAb against human ICAM-1 disclosed that in IPF, the expression of ICAM-1 was intensively enhanced on alveolar epithelial cells. These results suggest that ICAM-1 may contribute to the pathogenesis of IPF.

B-cell activation by membrane-bound antigens is facilitated by the interaction of VLA-4 with VCAM-1

VCAM-1 is one of the main ligands of VLA-4, an integrin that is highly expressed on the surface of mature B cells. Here we find that coexpression of VCAM-1 on an antigen-bearing membrane facilitates B-cell activation. Firstly, this is achieved by mediating B-cell tethering, which in turn increases the likelihood of a B cell to be activated. Secondly, VLA-4 synergizes with the B-cell receptor (BCR), providing B cells with tight adhesion and enhanced signalling. This dual role of VCAM-1 in promoting B-cell activation is predominantly effective when the affinity of the BCR for the antigen is low. In addition, we show that the VCAM-1 ectodomain alone is sufficient to carry out this function. However, it requires the transmembrane domain to segregate properly into a docking structure characteristic of the B-cell immunological synapse (IS). These results show that the VLA-4/VCAM-1 interaction during membrane antigen recognition enhances B-cell activation and this function appears to be independent of its final peripheral localization at the IS.

Inhibition of expression of P-selectin by antioxidant in cholesterol-fed rats.

Butylated hydroxytoluene (BHT) can inhibit experimental atherosclerosis in animals. Although the agent is an antioxidant, the exact mechanism of the reaction in atherosclerosis is still unknown. To investigate the effects of BHT on expression of P-selectin (PADGEM, GMP-140), intercellular adhesion molecule-1 (ICAM-1) and class II MHC (Ia) antigen, we proposed an experiment on rats. Male rats (n=18 per group) were fed either a normal cholesterol control diet, a normal cholesterol diet containing 0.5% BHT (BD), a high cholesterol diet containing 1.5% cholesterol and 0.1% sodium cholate (CD), or the CD diet containing 0.5% BHT (BCD). Rats were sacrificed after 3 days, and after 1, 2, 4, 10, and 17 weeks of dietary treatment. Although there was no gross or light microscopic atherosclerotic lesions, scanning electron microscopy revealed monocytic adhesion to aortic endothelium and mild endothelial injuries in CD and BCD groups. Immunohistochemically, the addition of BHT to a high cholesterol diet inhibited P-selectin expression but not in ICAM-1 and Ia antigen. These findings suggest that in rats, high cholesterol diets induce expression of ICAM-1, P-selectin and Ia antigen. In addition, the antiatherogenic effect of BHT may play a role in the inhibition of P-selectin.

Antiproliferative effect of calcitriol on human prostatic cancer cell lines: unrelated to the expression of major histocompatibility complex antigens or intercellular adhesion molecule-1 (ICAM-1).

In addition to its role in calcium and phosphorus metabolism, calcitriol (1,25-dihydroxyvitamin D3; 1,25-D3) demonstrates multiple effects on cell proliferation/differentiation by expressing major histocompatibility complex (MHC) antigens and intercellular adhesion molecule-1 (ICAM-1). It has recently been reported that 1,25-D3 inhibits the growth of prostatic cancer (PCa) cells. In this study we examined the effect of 1,25-D3 on both the growth and expression of HLA-ABC, HLA-DR and ICAM-1 antigens in PCa cells. Four human PCa cell lines (PC-3, PPC-1, ALVA-41 and ALVA-101) were examined. The cell numbers were enumerated, and the effects of interferon-y (IFN-gamma) and 1,25-D3 on the expression of HLA-ABC, HLA-DR and ICAM-1 were quantitated by flow cytometry. A dose-dependent antiproliferative effect of 1,25-D3 was found in all PCa cells lines except ALVA-41.1,25-D3 was approximately 10 times as potent as its analogue 24,25-dihydroxyvitamin D3 in inhibiting the growth of PC-3 cells. Also, the relative inhibitory ability of these compounds paralleled the strength of their binding affinities for the 1,25-D3 receptor, indicating that the antiproliferative effect may require a receptor-ligand interaction. HLA-ABC was expressed in PC-3, ALVA-41 and ALVA-101, but not in PPC-1 cells, while HLA-DR was not expressed on any of the tested cells. IFN-gammacould enhance or induce HLA-ABC but not HLA-DR expression in the tested cells. ICAM-1 was expressed in all cells and slightly upregulated by IFN-gamma. In this study 1,25-D3 had an antiproliferative effect on 3 of the 4 examined PCa cell lines.

Simultaneous Height and Adhesion Imaging of Antibody-Antigen Interactions by Atomic Force Microscopy

Specific molecular recognition events, detected by atomic force microscopy (AFM), so far lack the detailed topographical information that is usually observed in AFM. We have modified our AFM such that, in combination with a recently developed method to measure antibody-antigen recognition on the single molecular level (Hinterdorfer, P., W. Baumgartner, H. J. Gruber, K. Schilcher, and H. Schindler, Proc. Natl. Acad. Sci. USA 93:3477–3481 (1996)), it allows imaging of a submonolayer of intercellular adhesion molecule-1 (ICAM-1) in adhesion mode. We demonstrate that for the first time the resolution of the topographical image in adhesion mode is only limited by tip convolution and thus comparable to tapping mode images. This is demonstrated by imaging of individual ICAM-1 antigens in both the tapping mode and the adhesion mode. The contrast in the adhesion image that was measured simultaneously with the topography is caused by recognition between individual antibody-antigen pairs. By comparing the high-resolution height image with the adhesion image, it is possible to show that specific molecular recognition is highly correlated with topography. The stability of the improved microscope enabled imaging with forces as low as 100 pN and ultrafast scan speed of 22 force curves per second. The analysis of force curves showed that reproducible unbinding events on subsequent scan lines could be measured.

Effects of Retinoic Acid Combined with Irradiation on the Expression of Major Histocompatibility Complex Molecules and Adhesion/Costimulation Molecules ICAM-1 in Human Cervical Cancer

Objectives.Radiation treatment is one of the most standardized and effective modalities for contemporary cervical cancer therapy. In addition, the radiation-potentiating effects of retinoic acid have been recently described. In order to investigate whether enhanced immunogenicity might be responsible for such potentiation, we have evaluated the effects of retinoic acid combined with high doses of γ-irradiation on the expression of major histocompatibility complex (MHC) Class I and II and intercellular adhesion molecule-1 (ICAM-1) in human cervical carcinoma cell lines.Methods.The expression of surface antigens (MHC Class I and II and ICAM-1) was evaluated by FACS analysis in untreated control cells and in cells following their exposure to retinoic acid, high doses of γ-irradiation (i.e., 5000 and 10,000 cGy), or the combination of the two procedures.Results.HT-3 and SiHa cervical cancer cells expressed variable levels of MHC Class I and ICAM-1 antigens while Class II surface antigens were not detectable. Exposure to either 5000 or 10,000 cGy completely inhibited cell replication in both cell lines and significantly and consistently increased the expression of all surface antigens present on the cells prior to irradiation. Irradiation was unable to induce neoexpression of antigens previously not expressed by these cells (i.e., MHC Class II). In a similar fashion, retinoic acid was also able to significantly increase the expression of MHC Class I and ICAM-1 antigens when compared to untreated tumor cells but was not able to induce the expression of HLA Class II surface antigens. Exposure to the combination of radiation plus retinoic acid significantly upregulated HLA Class I and ICAM-1 molecules in an additive manner when compared to the levels obtainable with the exposure to radiation or retinoic acid alone.Conclusions.These data indicate that the combination of these two treatments could induce an additive effect on the expression of immunologically important surface antigens in human cervical cancer cells. These findings, together with the powerful antiproliferative effect of retinoids and irradiation on tumor cells, suggest that the combined regimen may be a promising and more effective combination for the treatment of cervical cancer.

Cytokine modulation of antigen expression in human melanoma cell lines derived from primary and metastatic tumour tissues.

The constitutive and cytokine-mediated expression of MHC class I and II antigens and intercellular adhesion molecule-1 (ICAM-1) was evaluated on eight human melanoma cell lines derived from primary and metastatic malignancies from patients (WM human melanoma series) including three pairs of related cell lines derived from the same individual. The cytokines IL-1 beta, IL-4, IL-6, TNF alpha, TGF beta 2, IFN gamma and IFN alpha were assessed for their ability to modulate the expression of cell surface antigens. MHC class I and class II antigen expression was unregulated by IFN gamma, IFN alpha and/or TNF alpha in cell lines established from primary melanoma. In contrast the cell lines derived from metastatic deposits did not show an increase in expression of MHC antigens in response to these cytokines. Both primary and metastatic WM cell lines were shown to be resistant to spontaneous natural killer cell (NK) activity, but susceptible to effector lymphocytes mediating lymphotine activated killer (LAK) cytotoxicity as a result of activation by IL-2. Although the constitutive and cytokine-induced level of expression of ICAM-1 and MHC antigens varied between paired primary and metastatic cell lines, this did not correlate with susceptibility of the cell line target to NK or LAK cytotoxicity. Whereas the IFNs, TNF alpha, TGF beta 2 and IL-1 beta differentially modulated the expression of ICAM-1 and MHC class I, treatment with IFNs (but not IL-1 beta, TNF alpha or TGF beta 2) resulted in a significant reduction in the sensitivity of the melanoma cells to NK and LAK cytotoxicity. Constitutive ICAM-1 expression was positively correlated with the ability of WM cell lines to colonise the lungs of SCID mice upon i.v. injection. The acquisition of cytokine resistance and inability to demonstrate enhanced cell surface expression may represent an important feature associated with the development of the metastatic phenotype.

Differential expression of immunoreactive HLA-DR and ICAM-1 in human cultured orbital fibroblasts and orbital tissue.

Intercellular adhesion molecule-1 (ICAM-1) and class II major histocompatibility complex antigens (MHC II, HLA-DR), are two cell-surface ligands known to be critical to the initiation and propagation of immune-mediated inflammation. Their expression in tissue is induced by proinflammatory mediators, principally interferon-gamma, tumor necrosis factor-alpha, and interleukin-1 beta. We examined human orbital fibroblasts and intact human orbital tissue exposed to these mediators for the presence of immunoreactive ICAM-1 and HLA-DR antigen. These ligands were detectable on stimulated fibroblasts and on fibroblasts and vascular endothelium in samples of orbital fat and muscle. ICAM-1 reactivity was diffuse and occurred in response to all three proinflammatory mediators. In contrast, HLA-DR reactivity was intense but limited to fewer numbers of cells. Our results, demonstrating the differential expression of these two ligands, may help further to elucidate the underlying pathogenetic mechanisms involved in the initiation and perpetuation of immune-mediated orbital inflammatory diseases.

Distribution of intercellular adhesion molecule-1 on leukocytes and corneal endothelium after endotoxin stimulation in rats

After stimulation with Salmonella typhimurium endotoxin, the intercellular adhesion molecule-1 (ICAM-1) was studied on the corneal endothelium and associated leukocytes in rats using immunoscanning electron microscopy. Two hundred micrograms of the endotoxin was injected in Lewis rats. The corneae were excised at 0-h and 16-h-postinjection time (n = 5, respectively). The corneae were prepared in hypothermic University of Wisconsin (UW) solution for immunoscanning electron microscopy. Histotopographical examination visualized ICAM-1 antigen on cytoplasmic processes of the corneal endothelium, arranged along microfolds, especially at the peaks. In the leukocytes, ICAM-1 was located primarily in morphologically non-specialized domains of the cell body surface, and only rarely scattered on the surface of microvillar projections. We concluded that the endotoxin stimulation can increase ICAM-1 in both corneal endothelium and associated leukocytes. Increased ICAM-1 may be an important factor for the leukocytes to form clustering and adhering to the corneal endothelium.

The phenotype of human eosinophils, basophils, and mast cells

Basophils, eosinophils, and mast cells (MCs) are specialized effector cells of the immune system. They are involved in inflammatory and allergic reactions and store and release multifunctional mediator molecules including vasoactive substances, lipid mediators, or cytokinesJ -3 Several different agonists, such as immunoglobulin, complement components, peptides, or cytokines and their specific receptors are involved in the activation of basophils, eosinophils, and MCs for mediator secretion? -~ MCs, basophils, and eosinophils originate from uncommitted multipotent progenitor cells. 5 Precursor cells giving rise to eosinophils and basophils (CFU eo/ba) are responsive to T cell-derived factors,6, 7 whereas MC precursor cells (CFUMAST) are responsive to stroma-cell-derived cytokines. 8 Basophils, eosinophils, and MCs form separate cell lineages and the committed precursor cells (giving rise to either MCs, basophils, or eosinophils) clearly are different cells?' 9 Basophils and eosinophils usually complete their differentiation in the bone marrow. After maturation both cells leave the marrow and are circulating cells. Under certain circumstances, and in particular on cell activation, basophils and eosinophils may leave the bloodstream to enter extravascular (inflamed) tissues. 3 In contrast, MCs derive from circulating multipotent progenitor cells (CFU-MIX), and after extravasation/homing of progenitors, undergo and complete their differentiation in extramedullary organs. 1° However, MCs may also migrate in the tissues and appear at sites of inflammation.

Interferon Gamma Induces Intercellular Adhesion Molecule-1 on Murine Midterm Trophoblast and Enhances Susceptibility to Specific Lysis by Paternally Directed Allo-Immune Cytotoxic T Cells1

Expression and function of intercellular adhesion molecule-1 (ICAM-1) on murine trophoblast cells and its regulation by interferon gamma (IFN-gamma) were investigated. Flow cytometry was used to detect ICAM-1 and class I major histocompatibility complex (MHC) antigen expression, while a 51Cr release assay was used to investigate the role of ICAM-1 in cytotoxic T lymphocyte (CTL)-mediated lysis and the effect of anti-ICAM-1 antibody blockade on lysis. We found that murine trophoblasts cells from midterm pregnancy (Day 14 postcoitum) express low or undetectable ICAM-1 and MHC antigens but that these are readily inducible by IFN-gamma. Untreated cells resisted lysis by allospecific CTL however, after treatment with IFN-gamma for 72 h, these trophoblasts were readily susceptible to lysis by allospecific CTL. The lysis was significantly reduced by anti-ICAM-1 antibody blocking. This finding which indicates that ICAM-1 can take part in CTL-mediated lysis of midterm trophoblast, has potentially important implications in vivo for the immunological relationship between mother and fetus.

Role of intercellular adhesion molecule-1 (ICAM-1) and lymphocyte function-associated antigen-1 (LFA-1) in peripheral blood mononuclear cell activation by human renal carcinoma cells.

We examined the role of intercellular adhesion molecule-1 (ICAM-1) and lymphocyte function-associated antigen-1 (LFA-1) during the activation of peripheral blood mononuclear cells (PBMCs) by the human renal carcinoma cell line CaKi-1. ICAM-1 antigen expression was induced on CaKi-1 cells by incubation with either phorbol-12-myristate 13 acetate (PMA) or interferon-gamma (IFN-gamma). Following a thorough washout of PMA and IFN-gamma and subsequent paraformaldehyde fixation, CaKi-1 cell monolayers were cocultered with allogenic PBMCs. While PMA-treated CaKi-1 cells induced PBMC proliferation and interleukin-2 receptor antigen expression, this was not the case for control or IFN-gamma-treated CaKi-1 cells. Furthermore, the induced PBMC proliferation was inhibited by specific monoclonal antibodies against ICAM-1 and LFA-1. Finally, although PMA induced human leukocyte antigen (HL)-A, B, C antigen expression on CaKi-1 cells, a monoclonal antibody against this antigen did not inhibit PBMC proliferation. We conclude that PMA can modulate CaKi-1 cells to stimulate allogenic PBMC proliferation in an ICAM-1/LFA-1 dependent, but HLA-A, B, C-independent, fashion. This stimulation might reside in the long-term activation of protein kinase C, induced by PMA.

Interleukin-10 Inhibition of HLA-DR Expression in Human Herpes Stromal Keratitis

Human herpes stromal keratitis (HSK) is an important cause of visual loss and morbidity. The presentation of corneal and/or viral antigens is thought to activate T lymphocytes, resulting in aberrant cell-mediated immune responses that are central to the pathogenesis of HSK. Aberrant cellular expression of HLA-DR and intercellular adhesion molecule-1 (ICAM-1), both of which are necessary for optimal antigen-induced T-lymphocyte responses, is present in lesions of HSK, but little is known concerning endogenous cytokines that may inhibit HLA-DR or ICAM-I expression in human disease. In this study, the authors investigated the effects of interleukin-10 (IL-10) on HLA-DR and ICAM-1 expression in human HSK. Penetrating keratoplasty specimens removed from 5 patients with HSK were divided to provide adjacent sections that were incubated with control medium or the same medium containing IL-10 (100 U/ml) for 48 hours. Immunoperoxidase staining was performed on each control and IL-10-treated corneal specimen to determine HLA-DR and ICAM-1 antigen expression. Interleukin-10 treatment resulted in profound reduction in immunoreactive HLA-DR, but not ICAM-1, in corneal cells and infiltrating leukocytes of all five HSK specimens. This study suggests that HLA-DR antigens may be selectively inhibited by cytokines released during inflammation in HSK. These results are the first to demonstrate cytokine suppression of HLA-DR in a human disease. Pharmacologic doses of IL-10 may inhibit HLA-DR-dependent immune responses that underlie a variety of destructive ocular inflammatory diseases.