Biomedical subjects
Publications and source records attributed to R P Laguens.
The expression of heat shock protein 27 (Hsp 27) in breast cancers correlates with stage of disease, the lower the stage the higher the expression, and with the presence or absence of lymph node metastases; lymph node negative patients being more likely to express Hsp 27 (P<0.04).
BACKGROUND: We studied whether the level of anti-skeletal muscle glycolipid antibodies (AGA), a marker of acute rejection in heart transplantation, may be associated with an adverse prognosis in unstable angina. METHODS AND RESULTS: The in-hospital evolution of 50 patients with unstable angina (Braunwald class III B) was assessed. We determined the incidence of death, myocardial infarction, and refractory angina. Blood was collected at admission and 24 hours later for determination of AGA levels by enzyme-linked immunosorbent assay. Twenty-three patients showed a decrease in the AGA level at 24 hours after admission. Ten in-hospital cardiac events occurred in these patients (43.4%) as compared with 4 (14.8%) in the 27 patients who did not show a decrease (P =.025). In patients with previous myocardial infarction (n = 26), the AGA assay was a powerful predictor of outcome. In this subgroup, 66.6% of patients who had decreased AGA levels (8 of 12) had cardiac events as compared with 14.2% (2 of 14) of those who did not have that decrease (P =.001). CONCLUSIONS: We conclude that a decrease of AGA levels 24 hours after admission is associated with a complicated in-hospital course. This finding may provide new insights in the phenomenon of plaque instability involved in the development of acute coronary syndromes.
Lymphocyte functional activity from lymph nodes draining human malignancies reflects the host immune response against tumour. Breast cancer is the neoplasia with the greatest amount of identified antigens but a weak inducer of a host efficient immune response. In our study we compared the mitogen stimulated-proliferative response of cells isolated from metastases-free lymph nodes draining breast cancer (Group 1), other malignant tumours (Group 2), and those obtained from patients without malignancies (Control group). A significant decrease of the proliferative response in cells isolated from lymph nodes draining breast cancer was observed comparing it to the other groups. Quantitative analysis of B and T cells showed a higher number of B cells than T cells in Groups 1 and 2. Moreover, Group 1 presented a two fold increase of T cells compared with Group 2. Our results suggest that the immunosuppression observed in lymph nodes draining breast cancer is higher than the inmunosuppression presented in other malignant tumours and that impaired function is not correlated with the increased number of T cells.
A rare development of acute inferior myocardial infarction is reported in a 23-year-old man with no previous history of cardiovascular disease. In an echocardiographic study a left intraventricular tumour was diagnosed. Cineangiographic study showed normal coronary arteries. The tumour, a myxoma, originating in the ventricular septum, was resected through the left atrium after the anterior leaflet of the mitral valve was detached. Postoperative course was uneventful and the patient remained healthy 48 months after surgery.
Studies carried out during the last decades provided evidence in support of an autoimmune pathogenesis for chronic chagasic myocarditis. This opinion was based on 1) the demonstration of molecular mimicry between parasite and host antigens, 2) the appearance of autoantibodies recognizing heart epitopes during the chronic phase of infection, 3) the induction of myocarditis and electrocardiographic alterations in animals immunized with whole parasites, parasite fragments or with biochemically-defined antigens, 4) the isolation from the heart of inflammatory infiltrates of B cells elaborating antibodies against myocardial antigens and 5) or of T cell clones reacting with heart epitopes and 6) induction of heart and nervous tissue alterations by transfer of lymphocytes from infected animals into naive syngeneic hosts. However, the characteristics of the inflammatory infiltrate in human myocarditis, displaying a wide variety of cells, many of them not involved in autoreactivity, such as the presence of giant cell granulomas and abundant eosinophils, as well as its focality and asynchrony, and the frequent association with pericarditis, casts doubts about the possibility of autoimmunity being responsible for the perpetuation of the myocarditis. This is supported by the recent observation that treatment of asymptomatic patients with trypanocidal drugs prevents the development of cardiopathy and that parasite components, either antigens or genomic fragments, are present at the site of the inflammatory lesions. On the basis of this new evidence, other alternative pathogenetic mechanisms should be sought to explain the appearance of a polymorphic long-lasting myocarditis that needs the presence of tiny fragments of parasites to develop. In addition to the well known immunological pathogenesis, the link between such a small amount of parasite components, below the level of microscopic detection, and the induction of such an extensive inflammatory infiltrate, represents interesting avenues for research in the near future.
In forty-five patients who underwent orthotopic heart transplantation, the titer of anti-human skeletal muscle glycolipid antibodies (AGA) present in the sera at the moment of transplantation was correlated with the number of histologically diagnosed cellular grade 3A and humoral acute rejection episodes during the first 120 days after transplantation. Determination of a cutoff value of 0.800 for the AGA level was determined by a receiver operating characteristic curve. Thirteen of 19 patients (68.4%) with an AGA titer above 0.800 developed 24 severe rejection episodes, and of the 26 patients with an AGA titer below 0.800, only 4 (15.3%) presented 6 severe rejection episodes during that time. This was especially evident for the humoral rejection episodes, which were diagnosed in only 1 of the 26 patients with AGA below 0.800 and in 7 of the 19 with AGA above 0.800. Comparison by univariate analysis of other well-known risk factors for a greater number of rejection episodes during the early posttransplant period with the AGA level at the moment of transplantation revealed that the latter distinguished a greater number of patients at risk than the other factors, such as a female donor, the lymphocyte direct cross-match, or the status of the patients at transplantation; the odds ratios were 6.33 for the AGA level, 3.17 for the direct cross-match, and 2.76 for the status at transplantation. By multiple logistic regression analysis, the only relevant risk factors in our group of patients were the AGA level (P=0.0009) and the status at transplantation (P=0.0285). These results indicate that determination of the AGA level at the moment of transplantation could represent a useful method for distinguishing which patients are at risk for a greater number of rejection episodes during the early posttransplant period, with a greater sensitivity than other risk factors.
Repair processes of cells submitted to an injury may exhibit an indirect identifying feature of its activity, based on the amount of connective tissue that could be traced with special stains and morphometry. Hydroxyproline concentration in tissue is also an index with which the presence and amount of repair mechanisms can be assessed. They may be quantitated appropriately in correlation with the end products of collagen, which represent the amount of cicatricial tissue. In order to correlate the size of the scar in myocardial infarcts with the amount of connective tissue, a determination of hydroxyproline levels after 30 days of acute experimental infarctions was performed in 3 groups of rats: 10 received a single dose of 2I U of human growth hormone (HGH) per week for 4 weeks, in a bolus injection immediately after the infarction was surgically induced; 10 received the same dose as above in daily injections during a week. Finally, another group of 10 animals which did not receive an active treatment served as controls. All animals were sacrificed at the end of a month. A morphometric study with Van Gieson stain was performed. Hydroxyproline levels were determined. Hydroxyproline levels were considerably lower in hGH-treated rats (T) compared to control (C) rats: 3.30 ug/mg +/- 0.1 vs. 4.10 +/- 0.29 (p < 0.05). Morphometry: T.: 5.23 +/- 2.45 vs. C.: 4.22 +/- 2.15 of the left ventricular wall. Three aneurysms were found in C group versus 1 in T. Human growth hormone administered to a group of rats with myocardial infarction showed obvious dose--effects consisting of a significant diminution of scar tissue in treated rats with a proportional fall in the hydroxyproline levels.
The mechanisms following intimal injury predisposing towards atherosclerotic changes have not been fully elucidated. We speculated that a local increase in the enzyme lipoprotein lipase (LPL) might explain a higher susceptibility of the damaged intima to lipid accretion, and so we investigated the effect of balloon endothelial denudation on LPL activity and cholesterol content (LPLa and Cholc, respectively), in aortas from normolipidemic male New Zealand white rabbits. Arteries were obtained from injured and control animals after 2, 6, 8 and 10 weeks to evaluate the shortest period after de-endothelialization necessary to detect LPLa changes. Injury resulted in a 4-fold LPLa rise (P < 0.01), as early as 2 weeks, and the enzymatic activity remained increased throughout the study period. A mild but significant 22% Cholc increase (P < 0.03) was found after 2 weeks of injury, even in this normolipidemic rabbit model. We conclude that physical damage to the intima markedly and soon increases LPLa. This finding might account for the higher lipid accumulation by injured vessels, providing additional support to the hypothesis of LPL as an atherogenic mediator.
In seventeen patients the result of the histological study of 153 endomyocardial biopsies (EMB) was compared with the ELISA titer of anti-human skeletal muscle glycolipid antibodies (AGA) present in serum samples collected simultaneously with the EMB procedure during the first four months following cardiac transplantation. The glycolipids were extracted from the quadriceps femoralis of blood group O patients. In the serum samples corresponding to the histological rejection grades with myocyte necrosis (greater than or equal to 2, International Society for Heart and Lung Transplantation grading) the AGA titer was significantly higher (P<0.005) than in the less severe rejection grades. The follow-up in each patient showed that the AGA titer raised in the serum samples collected immediately after, before, or coincidentally with a histological diagnosis of rejection grade 2 or 3A. In only one rejection grade 3A case was a false-negative result observed. Determination of the cut-off of the AGA level versus rejection grades 2 and 3A was determined by a relative-operating characteristic curve. An optical density (OD) of 0.040 showed maximum efficiency with sensitivity 53% and specificity 79%. Four patients who had AGA with an OD above 0.040 at the time of transplant had a significantly higher number of rejection grade 2 and 3A episodes than eleven patients with low pre-transplant AGA titers (P<0.05). These results indicate that search of anti-skeletal muscle glycolipid antibodies may represent a useful noninvasive method for monitoring heart rejection, and suggest that its investigation prior transplant may be a predictor of the number of grades 2 and 3A rejection episodes.
The immunization of biungulate animals with killed foot-and-mouth disease virus (FMDV) requires periodic vaccinations due to a low vaccine immunogenicity. Therefore, FMDV antigens need to be combined with adjuvants such as aluminium hydroxide, saponin or oil emulsions. Animal handling for periodic inoculations, and the repeated doses of vaccines that have to be administered increase the commercialization costs. Moreover, the use of adjuvants may induce adverse effects. In the present work we show that it is possible to increase the life span of neutralizing antibodies in serum when a single dose of cyclophosphamide (Cy) is administered four days before vaccination with aluminium hydroxidesaponin FMDV vaccine.
Administration of a low sub-immunosuppressive dose of cyclophosphamide (CY) to naive mice induced a marked increase in the number of splenic cells forming natural antibodies against unrelated antigens such as foot-and-mouth disease virus, keyhole limpet hemocyanin, horseradish peroxidase, or bovine serum albumin, as determined by an enzyme-linked immunosorbent assay spot technique. These results suggest that in mice there exists a repertoire of B cells forming natural antibodies which is restrained by an unknown mechanism and that CY is able to interfere with that restriction.
BACKGROUND AND METHODS: In sixty-three endomyocardial biopsy samples collected from six heart transplant recipients for the diagnosis of acute rejection episodes, the presence of apoptosis in individual cells was investigated in tissue sections by in situ labeling of nuclear DNA breaks by nick end labeling with biotinylated poly deoxyuridin triphosphate introduced by terminal deoxynucleotidyl transferase and alkaline phosphatase-conjugated streptavidin. In the samples collected at the moment of transplantation, no apoptotic cells were observed. Apoptotic nuclei were found in the myocytes and capillary endothelial and connective tissue cells of endomyocardial biopsy samples obtained from day 7 to day 146 after transplantation with a different prevalence according to the rejection grade (International Society for Heart and Lung Transplantation classification). RESULTS: In all the rejection grade 3A (eight of eight), in half of the rejection grade 2 (four of eight), and in some rejection grade 1B (three of eight) cases, apoptotic myocytes were found within or in the neighborhood of the inflammatory areas. In the rejection grades 0 and 1A and in the "Quilty" effect zones, no apoptotic myocytes could be observed. Apoptotic endothelial and interstitial cells were observed in all the rejection grades but with a higher prevalence in rejection grades 2 and 3A. CONCLUSIONS: During rejection episodes, apoptosis of myocytes is one of the mechanisms of immune-mediated death, and its investigation in tissue sections may represent a valuable tool for the diagnosis of myocyte damage.
Sixteen axillary lymph nodes were incubated with sera from patients with mammary carcinoma. Using immunofluorescence staining sera recognized antigenic determinants on follicular dendritic cells (FDC) within the follicle centers. These results were confirmed with isolated and cultured FDC that were incubated with the same sera. All the results were negative with normal sera. We also found a cell population positively reacting with a monoclonal antibody against an estrogen receptor associated protein (ERAP) in subcapsular and cortical sinusae and germinal centers. Phenotype identification of ERAP+ cells indicated that they presented characteristics of macrophages and FDC respectively. Lymph nodes from other malignancies were negative for ERAP. These findings suggest that the tumoral antigen could be either the protein associated with the estrogen receptor or the receptor itself. The ERAP could be transported by the macrophages from the tumor to the regional lymph nodes where it could be processed and maintained during a long time by FDC, since it is known that these are the most efficient antigen presenting cells.
OBJECTIVE: To characterize biochemically and isolate the skeletal and heart muscle cell epitope recognized by the autoantibodies present in the serum of chronically infected Trypanosoma cruzi patients. Secondly, to use that epitope in an immunoenzymatic assay for determining differences in antibody titre among Chagas' and other protozoan and heart diseases and between asymptomatic and cardiopathic chagasic patients. DESIGN: Isolated human skeletal and heart muscle cells were treated with organic solvents, pronase, neuraminidase and sodium metaperiodate before immunofluorescence assay. Glycolipids were extracted from human skeletal muscle for ELISA. PATIENTS: Sera were collected from 155 patients with positive serology for T cruzi infection; 44 healthy blood bank donors; and from patients after heart transplantation (16 patients), during the first month after cardiac infarction (eight) or cardiotomy (10), dilated myocardiopathy (21), leishmaniasis (12), acute toxoplasmosis (four) and hyperthyroid ophthalmopathy (five). MAIN RESULTS: Immunofluorescence assay revealed that the chagasic sera recognized epitopes that appeared to be glycolipid in nature. ELISA showed that the chagasic sera contained a higher titre of antiskeletal muscle glycolipid antibodies than the control sera and that, in the chagasic population, antibody titre was significantly higher in patients with heart failure than in asymptomatic subjects or in those presenting only electrocardiographic abnormalities. CONCLUSIONS: The skeletal and heart muscle epitope recognized by antibodies present in the sera of chagasic patients has the characteristics of a glycolipid. ELISA with glycolipids extracted from human skeletal muscle indicated that chagasic patients presented a higher antibody titre and that patients with heart failure showed a titre significantly higher than those who were asymptomatic or with electrocardiographic abnormalities, suggesting that those antibodies could be immunological markers and even predictors of heart failure in Chagas' disease.
Human and murine polymorphonuclear leukocytes (PMN) lysed L cells infected with Junin Virus (JV) in an in vitro system free of antiviral antibody and complement. Infected VERO cells proved to be resistant to the cytolytic effect. Murine PMN showed an increased adherence on JV-infected L cells but did not attach to VERO cells. The opposite was observed with human PMN, which did adhere to infected VERO cells but not to infected L cells. These results indicate that PMN activity may be an early mechanism of defense in JV infection by lysing virus-infected cells when no immune response has been established, and that cytoadherence is not a necessary step for target lysis.
In isolated skeletal, heart, and smooth muscle cells from BALB/c and C3H/HeJ mice infected with different strains of Trypanosoma cruzi the presence of class II MHC molecules was investigated by immunocytochemical techniques. We employed single muscle fibers instead of conventional cryostat sections to obtain a more accurate antigen localization. Approximately half of the skeletal muscle cells isolated from the rectus femoris expressed Ia antigens on their surface, irrespective of the mouse or parasite strain combination. Ia expression was apparent only at 30 days postinfection and thereafter. The heart muscle cells expressed class II molecules only at 1 and 3 months postinfection. In no case did the smooth muscle cells from infected mice express Ia antigens. Studies of the same molecules in the noninfected animals gave constantly negative results. We conclude that in the course of the chronic infection of mice with T. cruzi, ectopic expression of class II MHC molecules occurs at the surface of skeletal and heart muscle cells, providing a possible mechanism for explaining the anti-striped muscle autoreactivity present in Chagas' disease.
Chagasic myocarditis is associated with the appearance of circulating antiheart autoantibodies. In order to find out if there was local synthesis of those antibodies we investigated, by means of a solid immunoenzymatic technique, the presence of cells secreting antibody (ASC) against syngeneic soluble heart antigens in the mononuclear cell (MNC) population isolated from the hearts of mice chronically infected with Trypanosoma cruzi. In seven animals the number of ASC ranged between 300 and 2080 per 10(6) MNC. A similar number of cells was observed when the assay was carried out with T. cruzi-soluble antigens. When the ASC were enumerated in an assay simultaneously with both antigens, their number doubled that found in the single antigen assay, suggesting that there was no cross-reactivity between the heart and the parasite antigens. These results indicate that some of the cells in the inflammatory infiltrate of chronic chagasic myocarditis synthesize IgG autoantibodies against heart antigens, a phenomenon which may lead to a local concentration of antibody large enough to induce tissue damage.
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