Amarilla Veres

Relationship of Anti-60 kDa Heat Shock Protein and Anti-Cholesterol Antibodies to Cardiovascular Events

Background—Several recent studies have indicated an association between key inflammatory mediators and atherosclerotic diseases. We evaluated whether high levels of antibodies against heat shock proteins and cholesterol (ACHA) predicted cardiovascular (CV) events. Methods and Results—We used blood samples from the Heart Outcomes Prevention Evaluation (HOPE) study to conduct a nested case-control study of 386 cases with CV events and 386 age- and sex-matched HOPE study controls without events. We explored the relationship between anti-hsp antibodies, ACHA, and subsequent outcomes (incident myocardial infarction, stroke, or CV death) during a mean follow-up of 4.5 years using conditional logistic regression. High levels of anti-hsp65 antibodies (≥90th percentile) predicted CV events (OR, 2.1; 95% CI, 1.2 to 3.9, P =0.01). Anti-hsp60 antibodies did not predict any event type, whereas incident stroke developed significantly less frequently in patients with high ACHA levels. Anti-hsp antibodies and ACHA did not correlate with inflammatory (fibrinogen, C-reactive protein, interleukin-6, intracellular adhesion molecule-1) or infectious markers (C pneumoniae or cytomegalovirus antibodies). Anti-hsp65 antibodies (≥90th percentile) and fibrinogen (highest tertile) had a strong joint effect: patients with high concentrations of both had more CV events (OR, 5.5; 95% CI, 1.8 to 17.5, P =0.004) than patients with low levels of both. A similar joint effect (OR, 2.7; 95% CI, 1.3 to 5.7, P =0.01) was found for high levels of anti-hsp65 and presence of cytomegalovirus antibodies. Conclusions—Serum antibodies to hsp65 were associated with subsequent CV events in this study of high-risk patients, independent of conventional cardiovascular risk factors and other inflammatory markers.

Antibodies against the human heat shock protein hsp70 in patients with severe coronary artery disease.

Heat shock proteins (hsps) play complex role in the function of the immune system, they can activate both humoral and cellular immune response, as well the complement system. Although autoimmunity to hsp70 was implicated in certain autoimmune diseases and other conditions, the exact role of anti-hsp70 antibodies is not known. It was demonstrated by our previous work and others findings that antibodies against the 60 kDa hsps are strongly associated with coronary atherosclerosis and carotis disease. It is also known that there is increased hsp70 expression at different sites of atherosclerosis. Therefore our aim was to study whether level of anti-hsp70 antibodies correlate with the presence of severe coronary artery disease (CAD). We measured and compared anti-hsp70 IgG antibody levels in CAD patients (n=99) and healthy subjects (n=99) with ELISA. The frequency of these antibodies was high in both groups and there was no significant difference in the median level of anti-hsp70 antibodies between patients with severe CAD and controls (653 (400–1141) vs. 630 (326–1152) AU/mL, P=0.337). Adjustment for age, sex, BMI and lipid parameters did not change this result. Furthermore we did not find a correlation between anti-hsp70 antibody levels and certain risk factors of CAD (age, lipid parameters, body mass index, C-reactive protein, gender, smoking, diabetes and anti-hsp60 antibodies). By contrast, in accordance with our previous findings, anti-hsp60 and anti-hsp65 antibody levels were significantly higher in CAD patients, compared to this control group (p<0.0001 for both variables). We did not find any correlation between the levels of anti-hsp70 and anti-hsp60 or anti-hsp65 antibodies either in the patients or the controls. The exact role of hsp70 in atherosclerosis is controversial, but we suggest that humoral immunity against human hsp70 does not contribute to coronary atherosclerosis in contrast to antibodies against 60 kDa hsps.

Imaging lid-parallel conjunctival folds with OCT and comparing its grading with the slit lamp classification in dry eye patients and normal subjects

PURPOSE To visualize and describe the morphologic appearance of lid-parallel conjunctival folds (LIPCOFs) by using optical coherence tomography (OCT) and to relate it to dry eye signs and symptoms. METHODS The LIPCOF grade, noninvasive tear film breakup time (NIBUT), lipid layer interference pattern, and dry eye symptoms were recorded in 17 normal subjects and 33 patients with dry eye. LIPCOFs were evaluated with a slit lamp and visualized by OCT. Three different algorithms for OCT were developed to grade LIPCOFs according to tear meniscus height or the covering tear film on the folds. RESULTS The three OCT methods showed significant correlation with the slit lamp method (r = 0.470-0.473, P < 0.01). The OCT LIPCOF imaging methods were independent of NIBUT. The Dry Eye Questionnaire (DEQ) scores correlated with the height of the folds and the absence of tear film coverage of the folds (r = 0.574, P < 0.001 and r = -0.527, P < 0.001, respectively). The OCT LIPCOF grades correlated with the DEQ scores (r = 0.494, P < 0.001 and r = 0.310, P = 0.029, respectively). The slit lamp grade did not correlate with the DEQ scores in the whole population, but did in the normal group (r = 0.458, P = 0.024). The OCT LIPCOF grades showed inverse correlation with lipid pattern in the normal group (r = -0.422-0.481, P = 0.05); however, this association disappeared in the dry eye group. CONCLUSIONS OCT enabled a noninvasive, high-resolution method of imaging, evaluating, and classifying LIPCOFs. These new classifications correlated well with the slit lamp grade and the DEQ scores, promising a new, more objective evaluation of dry eye.

Epistatic effects of genes encoding immunoglobulin GM allotypes and interleukin-6 on the production of autoantibodies to 60- and 65-kDa heat-shock proteins

Immunoglobulin GM and KM genes have been associated with antibody responses to a variety of antigens. A promoter-region polymorphism of interleukin-6 (IL-6) gene (−174 G/C) has been shown to be associated with antibody responses to heat-shock proteins (hsp) 60 and hsp65. To examine the possible epistatic effects of these unlinked genetic systems on the autoimmune responses to hsp60 and hsp65, 176 healthy Caucasian subjects from Finland were genotyped for several allelic determinants of GM, KM, and IL-6 genes by PCR-RFLP methods. IgG antibodies to hsp60 and hsp65 were measured by an ELISA. Significant interactive effects of GM f,z and IL-6-174 genotypes were noted for both anti-hsp60 (P=0.002) and anti-hsp65 (P=0.038) antibody levels. Since these autoantibodies have been implicated in susceptibility to coronary heart disease and carotid atherosclerosis, the associations reported here might be relevant to the etiology of these diseases.

Impaired Humoral Immune Response Against Mycobacterial 65-kDa Heat Shock Protein (HSP65) in Patients with Inflammatory Bowel Disease

Since only scarce data are available on the immune response against heat shock proteins (HSP) in inflammatory bowel disease (IBD), we have measured with an ELISA method serum levels of IgG, IgA, and IgM antibodies to mycobacterial HSP65 and human HSP60 in 66 patients with Crohns disease (CD), 42 patients with ulcerative colitis (UC), and 126 age- and gender-matched healthy controls. Serum concentration [median (25th–75th percentiles) of IgG anti-HSP65 antibodies was substantially lower in patients with either CD (P < 0.01) or UC (P < 0.001) than in healthy controls, while no difference was found in the levels of anti-HSP60 antibodies. Low anti-HSP65 antibody levels were measured in patients with active CD and in both active and inactive UC, and only in IBD patients with no extraintestinal manifestations. In conclusion, our present findings indicate that an abnormal immune response to bacterial HSP65 or some epitopes of the protein may contribute to the dysregulation of host defenses against certain intestinal bacteria.

Complement activation in the nasal mucosa following nasal ragweed-allergen challenge

The aim of this study was to explore complement activation in the nasal lavage following a nasal ragweed‐allergen challenge. The study was carried out with 15 adolescents who were allergic to ragweed and with six non‐allergic healthy volunteers. Following the baseline measurement after the symptoms were registered, subjects were given increasing doses of ragweed allergen. Lavage fluid was collected and tested for a complement‐activation product (C3bBbP). The allergic patients responded to allergen provocation with an increase in C3bBbP formation compared to the initial lavage (p = 0.001). The C3bBbP level remained low in the lavage fluids of the non‐allergic controls. We found a strong correlation between the threshold dose that induced symptoms and the dose where the maximum complement activation was detected (r = 0.78, p = 0.001). Our findings indicate that in allergic patients nasal challenge with ragweed allergen induces a rise in complement activation in the nasal lavage fluid. These results highlight the role of the complement system in the allergic inflammation on the nasal mucosal surface.

C1r-C1s-C1inhibitor (C1rs-C1inh) complex measurements in tears of patients before and after penetrating keratoplasty

Purpose. The aim of this pilot study was to determine the presence of complement activation products in tears from pre- and postkeratoplasty eyes and the fellow eyes in order to investigate the activation of the classical and alternative pathways of the complement system in the early postkeratoplasty period. Methods. Tear samples from both eyes of 19 prekeratoplasty patients were tested. From 10 patients, samples were taken before operation, one week and 3 weeks after penetrating keratoplasty. Only baseline and 1 weak samples, and baseline and 3 week samples were available from 5 and 2 patients, respectively, while only baseline tear samples were collected from 2 patients. Tear concentration of two complement activation products, C1rs-C1inh and C3bBbP were determined by enzyme-linked immunosorbent assay. Results. There was no difference (p = 0.339) between baseline samples of the eyes waiting for operation (0.93 ± 0.51 AU/ml, mean ± SEM) and the fellow eyes (0.33 ± 0.33 AU/ml) in respect of mean C1rs-C1inh complex concentration. The one-week samples of the operated eyes revealed significantly (p = 0.006) elevated levels of C1rs-C1inh complexes (18.8. ± 6.37 AU/ml), compared to their baseline samples (1.18 ± 0.64 AU/ml), whereas the one-week values of the fellow eyes did not differ from the baseline values. Compared to the increased one-week values, the three-week values decreased to the baseline values in the operated eyes. C3bBbP could be detected in 3/68 tear samples. Conclusions. In our study we demonstrated the increased concentration of C1rs-C1inh complex in several tear samples taken early after human penetrating keratoplasty. These findings provide direct evidence that the classical pathway of complement may be activated in the early postoperative period after penetrating keratoplasty.

Tear film function in patients with seasonal allergic conjunctivitis outside the pollen season.

Background: Seasonal allergic conjunctivitis can manifest itself through tear film instability and symptoms of eye discomfort during the pollen season. This study investigated whether seasonal allergic inflammation defines tear film instability outside the season. Methods: Twenty-three control subjects and 13 ragweed-allergic patients were involved (21 female, 15 male; mean age 26.6 ± 5.4 years). Outside the pollen season, subjective ocular symptoms, non-invasive tear film break-up time, lower tear meniscus height and the tear lipid layer’s interference pattern grade were recorded. C3a complement activation level was also measured in collected tear samples. Results: Non-invasive tear film break-up time, lower tear meniscus height, C3a complement activation level and the incidence of the different grades of tear lipid pattern did not differ significantly in the two examined groups (p ≧ 0.223). The mean eye symptom score outside the season was greater in the allergic group, but the difference was not significant (p = 0.062). The C3a complement activation level showed a significant and inverse correlation with the lipid layer grade (r = –0.343, p = 0.017). Among the participants with thinner tear lipid layers, the complement activation in the tear samples was higher than among those patients with normal tear lipid layers. Conclusion: Seasonal allergic inflammation did not cause permanent tear film instability and eye symptoms were not observed outside the pollen season.

Kinetic analysis of topographical parameters and interference pattern of tear lipid layer in normal subjects

Purpose:  To investigate the post‐blink changes of the topographical regularity indices, the corneal wavefront aberrations and the tear lipid layer’s interference pattern, and analyze the relationship between these parameters’ changes.