Β2-microglobulin Expression Research Articles

HLA-DR and β2 microglobulin expression in medullary and atypical medullary carcinoma of the breast: histopathologically similar but biologically distinct entities

Aims—To examine the expression of HLA-DR and β2 microglobulin in medullary carcinoma and atypical medullary carcinoma of the breast to determine if the effective presentation of tumour antigens to the...

Recognition of a Shared Human Prostate Cancer-Associated Antigen by Nonclassical MHC-Restricted CD8+ T Cells

To identify prostate cancer-associated Ags, tumor-reactive T lymphocytes were generated using iterative stimulations of PBMC from a prostate cancer patient with an autologous IFN-gamma-treated carcinoma cell line in the presence of IL-2. A CD8+ T cell line and TCR alphabeta+ T cell clone were isolated that secreted IFN-gamma and TNF-alpha in response to autologous prostate cancer cells but not to autologous fibroblasts or lymphoblastoid cells. However, these T cells recognized several normal and malignant prostate epithelial cell lines without evidence of shared classical HLA molecules. The T cell line and clone also recognized colon cancers, but not melanomas, sarcomas, or lymphomas, suggesting recognition of a shared epithelium-associated Ag presented by nonclassical MHC or MHC-like molecules. Although Ag recognition by T cells was inhibited by mAb against CD8 and the TCR complex (anti-TCR alphabeta, CD3, Vbeta12), it was not inhibited by mAb directed against MHC class Ia or MHC class II molecules. Neither target expression of CD1 molecules nor HLA-G correlated with T cell recognition, but beta2-microglobulin expression was essential. Ag expression was diminished by brefeldin A, lactacystin, and cycloheximide, but not by chloroquine, consistent with an endogenous/cytosolic Ag processed through the classical class I pathway. These results suggest that prostate cancer and colon cancer cells can process and present a shared peptidic Ag to TCR alphabeta+ T cells via a nonclassical MHC I-like molecule yet to be defined.

Regulation of β2-microglobulin expression in different human cell lines by proinflammatory cytokines

Proinflammatory monocytic cytokines such as interleukin-1 (IL-1), tumour necrosis factor-alpha (TNF-alpha) and IL-6 have been incriminated in the pathogenesis of elevated beta2-microglobulin (beta2M) serum concentrations in patients undergoing haemodialysis with so-called bioincompatible dialyser membranes. However, neither the source of the elevated serum beta2M nor the precise role of monocytic cytokines in the expression of the beta2M gene have been elucidated conclusively. The aim of the current study was to evaluate whether monocytic cytokines, and in particular IL-6, are regulators of beta2M gene expression in human hepatoma cells, T-lymphocytes and monocytes. HepG2 and HuH7 human hepatoma cells, Jurkat T-cells, monocytic MonoMac6 cells, primary human monocytes and synoviocytes were stimulated with IL-1beta, IL-6, interferon-alpha (IFN-alpha), IFN-gamma or conditioned media from lipopolysaccharide (LPS)-treated monocytes. Expression of beta2M mRNA was analysed by Northern blotting, beta2M protein synthesis was determined by metabolic labelling and immunoprecipitation, and beta2M secretion was measured by an enzyme-linked immunosorbent assay (ELISA). In all cell types tested, IFN-gamma and, to a lesser extent, IFN-alpha stimulated gene expression of beta2M resulting in an increased synthesis and secretion of beta2M protein. Neither IL-1beta and IL-6 nor supernatants from LPS-treated monocytes were capable of inducing beta2M gene expression, with the exception of a small increase in HuH7 hepatoma cells upon IL-1beta treatment. The present study provides evidence that interferons are important regulators of beta2M expression. It also shows that proinflammatory monocytic cytokines do not modulate directly the expression of beta2M in cells of hepatic, monocytic and T-lymphocytic origin. Whether they influence beta2M synthesis and secretion indirectly by modulating interferon synthesis needs to be elucidated.

Differing MHC class I requirements for induction and propagation of experimental systemic lupus erythematosus

Mice deficient in β2-microglobulin expression are resistant to the induction of experimental systemic lupus erythematosus (SLE). The present studies were designed to identify the β2-microglobulin-dependent cell surface molecule(s) that confers sensitivity to experimental SLE, and to determine its role in disease development. We report hat mice lacking the transporter associated with antigen presentation (TAP–/–) were also resistant to disease, whereas CD1–/– and CD8–/– mice were susceptible; susceptibility also did not correlate with neonatal Fc receptor or HEPH expression. These data indicate that disease susceptibility is determined by expression of MHC class I. Furthermore, by analyzing both adoptive transfer and radiation bone marrow chimeras, we demonstrate that MHC class I expression is necessary for propagation of disease, but not for induction of pathogenic cells.

An increased number of follicles containing activated CD69+ helper T cells and proliferating CD71+ B cells are found in H. pylori-infected gastric mucosa.

An immune response occurring in Helicobacter pylori-infected gastric mucosa could have a direct implication for associated pathologies. In the present study we analyzed the expression of the immune activation, proliferation, and phenotype markers by immune cell subpopulations in H. pylori-infected and uninfected gastric samples. Antral gastric biopsies from both H. pylori-positive and -negative patients were processed by immunohistochemistry; gastric epithelial cells were isolated from biopsy tissue and analyzed by flow cytometry. Ten of the 13 biopsies that contained follicles were H. pylori positive. Follicular CD69 was expressed mainly by CD4+ T cells and the central core of follicles showed double immunopositivity for the B-cell marker CD19 and transferrin receptor. Also detected was an increase in the percentage of epithelial cells from H. pylori-positive samples expressing HLA-DR and beta2 microglobulin, compared to negative samples (61 +/- 15% vs 9 +/- 9%, p = 0.003 and 93 +/- 7% vs 52 +/- 18%, p = 0.002, respectively), whereas no variation on class I HLA was detected. These results suggest that chronic H. pylori infection could facilitate the persistence of follicles on which continuous follicular helper T-cell activation could lead to uncontrolled follicular B-cell proliferation. Furthermore, beta2 microglobulin expression by epithelial cells in a nonparallel way to class I HLA may indicate the possibility of nonclassical class I MHC expression on the basal surface of the epithelium.

An Increased Number of Follicles Containing Activated CD69 Helper T Cells and Proliferating CD71 B Cells Are Found in H. pylori-Infected Gastric Mucosa

Objective: An immune response occurring in Helicobacter pylori-infected gastric mucosa could have a direct implication for associated pathologies. In the present study we analyzed the expression of the immune activation, proliferation, and phenotype markers by immune cell subpopulations in H. pylori-infected and uninfected gastric samples. Methods: Antral gastric biopsies from both H. pylori-positive and -negative patients were processed by immunohistochemistry; gastric epithelial cells were isolated from biopsy tissue and analyzed by flow cytometry. Results: Ten of the 13 biopsies that contained follicles were H. pylori positive. Follicular CD69 was expressed mainly by CD4+ T cells and the central core of follicles showed double immunopositivity for the B-cell marker CD19 and transferrin receptor. Also detected was an increase in the percentage of epithelial cells from H. pylori-positive samples expressing HLA-DR and β2 microglobulin, compared to negative samples (61 ± 15% vs 9 ± 9%, p = 0.003 and 93 ± 7% vs 52 ± 18%, p = 0.002, respectively), whereas no variation on class I HLA was detected. Conclusions: These results suggest that chronic H. pylori infection could facilitate the persistance of follicles on which continuous follicular helper T-cell activation could lead to uncontrolled follicular B-cell proliferation. Furthermore, β2 microglobulin expression by epithelial cells in a nonparallel way to class I HLA may indicate the possibility of nonclassical class I MHC expression on the basal surface of the epithelium.

Deglucosylation of N-linked glycans is an important step in the dissociation of calreticulin-class I-TAP complexes.

Recent evidence indicates that newly synthesized major histocompatibility complex (MHC) class I proteins interact with calnexin, a transmembrane endoplasmic reticulum protein specific for certain glycoproteins bearing monoglucosylated glycans. Here, we studied the association of newly synthesized class I proteins with calreticulin, a soluble calnexin-related ER protein, in murine T cells. We found that, unlike calnexin-class I interactions, calreticulin assembly with class I proteins was markedly decreased in the absence of beta 2 microglobulin expression and that calreticulin associated with a subset of class I glycoforms distinct from those assembled with calnexin but similar to those bound to TAP (transporter associated with antigen processing) proteins. Finally, these studies show that deglucosylation of N-linked glycans is important for dissociation of class I proteins from both calreticulin and TAP and that the vast majority of newly synthesized class I proteins associated with calreticulin are simultaneously assembled with TAP. The data demonstrate that calnexin and calreticulin chaperones assemble with distinct MHC class I assembly intermediates in the ER and show that glycan processing is functionally coupled to release of MHC class I proteins from peptide transport molecules.

Expression of β2-microglobulin by premalignant epithelium

Many human tumors express low amounts of HLA class I molecules relative to the normal cells from which they are derived. From experimental work it is clear that the malignant behavior of a tumor cell may depend on its MHC class I expression. Therefore, it is of obvious interest to study the HLA class I expression of human tumors in their various stages. We have studied the HLA class I expression by the cells in premalignant epithelial lesions and invasive carcinoma of the bladder and uterine cervix using immunoperoxidase staining for beta 2-microglobulin of paraffin-embedded tissue. We here assume that beta 2-microglobulin expression by malignant and premalignant cells equals HLA class I expression. Thirty-two of the 36 invasive tumors expressed less overall beta 2-microglobulin than cells from the normal epithelium. In contrast, approximately two-thirds of 34 premalignant bladder epithelia and 47 premalignant cervix epithelia displayed higher overall beta 2-microglobulin expression than the normal epithelium. Thus, a systematic large-scale elimination of HLA class I high-expressing tumor cell variants may take place only after the tumor penetrates the basement membrane.

Expression of Major Histocompatibility Complex Antigens and Serum Neutralizing Antibody in Murine Retroviral Encephalitis

Murine leukemia virus infection serves as a model for noninflammatory degeneration of the central nervous system (CNS). During the course of infection with either of the molecularly cloned viruses pNE-8 or ts-1, we observed that ts-1 spread twice as rapidly as pNE-8, and ascended higher in the neuraxis. Endothelial cells were infected first, followed by oligodendrocytes and neurons, while astrocytes containing glial fibrillary acidic protein were not infected. Additionally, ts-1 also infected macrophages/microglia. Major histocompatibility complex (MHC) class I beta 2-microglobulin expression was minimal in pNE-8 infected mice, while it was elevated in endothelial cells of early ts-1 lesions, and in macrophages/microglia during later stages. Occasional infected cells expressed beta 2-microglobulin while rare endothelial and parenchymal cells expressed MHC class II in both viral infections. Limited intra-CNS MHC expression may be one of the mechanisms of viral persistence and will present a barrier to developing immunotherapy for CNS retroviral infections. The few mice that escaped lethal infection had higher serum titers of neutralizing antibodies and showed no neuropathologic changes or detectable virus in the CNS. Higher titers of neutralizing antibodies may protect the CNS from infection.