Glomerular expression and elevated serum Bcl-2 and Fas proteins in lupus nephritis: preliminary findings.

Programmed cell death (apoptosis) is involved in glomerular injuries leading to glomerulonephritis. Bcl-2 and Fas are proteins that promote cell survival and death, respectively. This study tests the hypothesis that lupus nephritis is associated with alterations of Bcl-2 and Fas protein expression. Thirty-six patients with lupus nephritis and 10 controls (normal individuals) were included in this study. Bcl-2 and Fas positive cells were examined in kidney biopsies by immunohistochemistry. Bcl-2 and Fas serum levels were evaluated by enzyme-linked immunosorbent assay (ELISA). In the glomeruli of normal kidneys, Bcl-2 and Fas proteins were completely absent. In lupus nephritis patients, glomerular expression of Bcl-2 and Fas was seen in mesangial cells (1.3 +/- 0.1 and 2.0 +/- 0.1 for Bcl-2 and Fas, respectively). Similarly, a statistically significantly higher Bcl-2 (217.1 +/- 85.9) and Fas (767.9 +/- 271) serum levels were found in lupus patients compared to controls (148.6 +/- 87, 550.3 +/- 91 for Bcl-2 and Fas, P < 0.05). A direct correlation between serum Bcl-2 and Fas and chronicity index was also found. Compared to normal controls, lupus nephritis is associated with glomerular expression and elevated serum levels of Bcl-2 and Fas proteins. These findings suggest possible roles for Bcl-2 and Fas in glomerular injury during evolution of lupus nephritis. The diagnostic, prognostic and therapeutic ramifications of our findings are open to further investigation.

Analysis of the mononuclear inflammatory cell infiltrate in the normal breast, benign proliferative breast disease, in situ and infiltrating ductal breast carcinomas: preliminary observations.

BACKGROUND: Mammary carcinogenesis is a multistep process entailing the transition from normal breast to benign proliferative breast disease (ductal hyperplasia) to ductal carcinoma in situ to infiltrating ductal carcinoma. HYPOTHESIS: These transitions are associated with changes in the mononuclear inflammatory cell infiltrate. MATERIALS AND METHODS: A total of 53 mastectomy specimens of normal breast, benign proliferative breast disease, ductal carcinoma in situ and infiltrating ductal carcinoma were evaluated for mononuclear inflammatory cell infiltrate by using immunohistological methods and monoclonal antibodies including CD20, CD68, CD3 and granzyme B, histiocytes, T cells and cytotoxic T cells. RESULTS: Transitions from normal breast to the subsequent tissue with lesions (normal skin v benign proliferative breast disease v ductal carcinoma in situ v infiltrating ductal carcinoma) were associated with significantly (p<0.01) increased mean (SD) density of mononuclear inflammatory cell infiltrate at the parenchyma (3.2 (1.0) v 26.4 (7.8) v 33.6 (7.9) v 39.1 (4.7) for CD20+ B cells; 2.8 (1.0) v 81.5 (14.0) v 84.0 (14.9) v103.7 (3.9) for CD3; 1.3 (2.0) v 3.8 (4.0) v 12.7 (23) v 22.1 (25.0) for CD68+ macrophages; 2.0 (1.0) v 58.3 (5.0) v 60.0 (10.0) v 74.1 (28.0) for granzyme B+ cytotoxic T cells) and at the stroma (0.7 (1.0) v 3.0 (5.0) v 13.3 (20) v 16.7 (30.0) for CD20+ B cells; 1.0 (2.06) v 4.0 (2.5) v 16.7 (5.0) v 21.7 (15) for CD68+ macrophages; 1.4 (0.6) v 4.2 (1.2) v 46.6 (16.7) v 77.0 (5.0) for CD3+ cells and 0 (0) v 0.5 (1.0) v 0.7 (1.0) v 0.7 (1.0) for granzyme B+ cytotoxic T cells). CONCLUSIONS: The increased mononuclear inflammatory cell infiltrate during mammary carcinogenesis may reflect non-specific or specific immunological processes.

Alterations of mononuclear inflammatory cells, CD4/CD8+ T cells, interleukin 1beta, and tumour necrosis factor alpha in the bronchoalveolar lavage fluid, peripheral blood, and skin of patients with systemic sclerosis.

BACKGROUND: Systemic sclerosis (SSc) is a multisystem disease with underlying immune mechanisms. AIMS: To investigate the clinicopathological characteristics of the lesions; immunological alterations in the bronchoalveolar lavage fluid (BALF), peripheral blood, and skin; and correlations between the clinicopathological characteristics and immunological alterations in SSc. MATERIALS/METHODS: Skin biopsies, BALF, and peripheral blood samples were obtained from 19 patients (18 women, one man) with SSc and six age and sex matched healthy controls (HCs). Mononuclear inflammatory cells (MICs), CD4/CD8 cells, tumour necrosis factor alpha (TNFalpha), and interleukin 1beta (IL1-1beta) concentrations were examined in all samples using histological methods, enzyme linked immunosorbent assay, and immunoperoxidase staining. RESULTS: The mean (SD) age of the patients with SSc was 34.8 (2.6) years. Proteinuria, positive rheumatoid factor, and C reactive protein were seen in 15.8%, 26.3%, and 26.3% of patients, respectively. Compared with HCs, there were significantly higher: total MICs (macrophages, lymphocytes), neutrophils, and eosinophils in BALF, blood, and skin (all p<0.05); cytokine concentrations in BALF (TNFalpha, p<0.001; IL-1, p<0.01) and peripheral blood (p<0.01 and p<0.05); and CD8/CD4+ T cells in peripheral blood (p<0.05). Compared with HCs, lesional skin had significantly higher histiocyte cell counts (p<0.05), lower lymphocyte counts (p<0.05), and higher CD4/CD8 ratios (p<0.001). There were significant correlations between cytokine concentrations and CD8+ T cells and forced vital capacity (p<0.001 and p<0.01, respectively). CONCLUSIONS: MICs, CD4/CD8+ cells, and cytokines are altered in SSc. These alterations correlated with the underlying disease process and therefore may have pathogenic, modulatory, and potential prognostic roles in SSc.

Altered expression of epidermal growth factor receptor in non-involved tissue of cancer-containing breasts.

Previous studies have identified functional differences in non-involved breast tissue from cancer-containing breasts. This study has examined the expression of epidermal growth factor (EGFR) protein and mRNA in the non-involved breast of over 100 cancer-containing breasts and compared these with the same number of normal breast tissues from age-matched women with no history of breast cancer. Immunohistochemistry with EGFR1 antibody applied to frozen sections was used for the detection of protein, and in-situ hybridization using a digoxigenin-labelled oligonucleotide probe was the method for detecting mRNA. EGFR protein was detected in myoepithelial cells and to a lesser extent in epithelial cells, where it was predominantly basal or baso-lateral. There was a significant difference in the extent of staining in ducts and lobules between non-involved tissue from cancer-containing breasts and age-matched normal breasts, it being significantly greater in the latter (P<0.001). Labelling for EGFR mRNA was greater and more consistent in myoepithelial cells than epithelial cells overall. Differences were found for intensity of labeling, with it being significantly greater for normal breast tissue (P<0.001) than non-involved tissue from cancer-containing breasts. There is reduced EGFR expression in normal breast tissue from cancer-containing breasts when compared to age-matched breast tissue from women with no history of breast cancer. The mechanisms underlying this are unclear but in previous studies we have identified alterations in myoepithelial cells in cancer-containing breasts and the present findings may represent altered myoepithelial cell function.

Decreased apoptosis in non-involved tissue from cancer-containing breasts.

Various histological changes in the breast have been associated with an increased risk of development of breast cancer; these changes have been identified in non-involved tissue in cancer-containing breasts, suggesting that factors promoting the development of carcinoma may have a field effect. Previous work has identified alterations in growth factors receptors and integrins in cancer-containing breast tissue. In the present study, proliferation and apoptosis are examined. Non-involved breast tissue from 104 women taken at least 4 cm away from a carcinoma and normal/benign tissue from 105 age-matched women were studied. Proliferation was assessed using MIB-1 immunohistochemistry and labelling for histone mRNA, as a marker of S-phase. In situ end-labelling was used to identify apoptosis; any non-labelled apoptotic bodies were also counted. No differences were found between the non-involved tissues and the control group for MIB-1 index and histone index. The apoptotic index was higher in the control group than in the cancer-containing breasts, being greater for ducts than for acini. When the apoptotic index/MIB-1 index and apoptotic index/histone index were considered, the mean for both was lower in the acini from cancer-containing breasts than in the control group, although the ratios for ducts were similar. The reduction in apoptosis may lead to the preservation of genetically aberrant cells, hence favouring neoplastic development. There is a need for further investigation of 'at-risk' cases, including women with a family history, and for a prospective study of a large group of women.