Postinfarction Cardiac Remodeling Proceeds Normally in Granulocyte Colony-Stimulating Factor Knockout Mice.

Treatment with granulocyte colony-stimulating factor (G-CSF) reportedly mitigates postinfarction cardiac remodeling and dysfunction. We herein examined the effects of G-CSF knockout (G-CSF-KO) on the postinfarction remodeling process in the hearts of mice. Unexpectedly, the acute infarct size 24 hours after ligation was similar in the two groups. At the chronic stage (4 weeks later), there was no difference in the left ventricular dimension, left ventricular function, or histological findings, including vascular density, between the two groups. In addition, expression of vascular endothelial growth factor (VEGF) was markedly up-regulated in hearts from G-CSF-KO mice, compared with wild-type mice. Microarray failed in detecting up-regulation of VEGF mRNA, whereas G-CSF administration significantly decreased myocardial VEGF expression in mice, indicating that G-CSF post-transcriptionally down-regulates VEGF expression. When G-CSF-KO mice were treated with an anti-VEGF antibody (bevacizumab), cardiac remodeling was significantly aggravated, with thinning of the infarct wall and reduction of the cellular component, including blood vessels. In the granulation tissue of bevacizumab-treated hearts 4 days after infarction, vascular development was scarce, with reduced cell proliferation and increased apoptosis, which likely contributed to the infarct wall thinning and the resultant increase in wall stress and cardiac remodeling at the chronic stage. In conclusion, overexpression of VEGF may compensate for the G-CSF deficit through preservation of cellular components, including blood vessels, in the postinfarction heart.

Identification of target antigens of antiendothelial cell antibodies against human brain microvascular endothelial cells in healthy subjects.

Antiendothelial cell antibodies (AECAs) have been detected in patients who have autoimmune and inflammatory diseases. Previous studies showed that AECAs against human umbilical vein endothelial cells were detected in healthy subjects. In the present study, we evaluated AECAs against human brain microvascular endothelial cells (HBMEC) in serum. We detected 250 antigen spots that reacted with AECAs in serum samples from 30 healthy subjects by 2-dimensional immunoblot using primary cultured HBMEC as the antigen source. There were 10 spots that corresponded to common target antigen spots and reacted with AECAs in serum samples from > 25% of the 30 healthy subjects. We identified 7 proteins that corresponded to 8 of the 10 spots by mass spectrometry: 78-kDa glucose-regulated protein, dihydropyrimidinase-related protein 2, heterogeneous nuclear ribonucleoprotein L, vimentin, perilipin 3, alpha-enolase, and annexin A2. The results suggest that these 7 HBMEC proteins are major target antigens of natural AECAs.

Anti-endothelial cell antibodies in patients with cerebral small vessel disease.

The pathogenesis of cerebral small vessel disease (CSVD) in the elderly is poorly understood. Endothelial cell activation and dysfunction may play a causal role in the pathogenesis of CSVD. It was reported that anti-endothelial cell antibodies (AECAs) are associated with endothelial cell dysfunction and inflammation. We hypothesized that AECAs may be associated with the pathogenesis of CSVD. We examined AECAs in sera from 12 elderly subjects with CSVD, 12 elderly subjects without CSVD, and 18 healthy volunteers by 2-dimensional immunoblotting using primary cultured human brain microvascular endothelial cells as the antigen source. We identified 4 AECAs that were detected in sera from more than one-half of the elderly subjects with CSVD. Subsequently, we analyzed the target antigens of these 4 antibodies by liquid chromatography-tandem mass spectrometry. The target antigens of these 4 antibodies were tropomyosin alpha-4 chain (TPM4), vimentin, alpha-enolase, and annexin A2. Among these 4 antibodies, the anti-TPM4 antibody was significantly more frequently detected in sera from the elderly subjects with CSVD than the other subjects. We determined the anti-TPM4 antibody level in sera from 21 elderly subjects with CSVD and 25 subjects without CSVD by enzyme-linked immunosorbent assay. The anti-TPM4 antibody level was significantly higher in the subjects with than without CSVD. Therefore, an autoimmune, inflammatory process with high levels of anti-TPM4 antibody may contribute to the development of CSVD in the elderly.

Fatal delayed hemolytic transfusion reaction associated with anti-Di(b) and anti-E.

We report the death of a 61-year-old Japanese man massively transfused during and after emergency aortic surgery. Postoperative on day 8, he died after cardiac arrest associated with hyperkalemia. Indirect antiglobulin testing demonstrated both anti-Di(b) and anti-E antibodies pre-transfusion, and elevation of their titers as the delayed hemolytic transfusion reaction evolved. Monocyte monolayer assay (induction of reactive monocytes) and flow cytometry (increase of IgG1 and/or IgG3) gave evidence of the clinical significance of both antibodies. Anti-Di(b) must be considered when an antibody to a high incidence antigen is found in Japanese and other Mongoloid populations.

Antibodies against the tom40 subunit of the translocase of the outer mitochondrial membrane complex and cognitive impairment in Alzheimer's disease.

Recent studies suggest that microvascular abnormalities are involved in pathology and progression of Alzheimer's disease. The purpose of this study was to examine the presence of antibodies against cerebral microvascular endothelial cells specific for Alzheimer's disease, and to evaluate the association of these antibodies with cognitive impairment. The study included patients with Alzheimer's disease (age ≥60 years; 24 patients), control subjects without neurological diseases (age ≥60 years; 19 subjects), patients with multiple sclerosis (all ages; 17 patients), and healthy control subjects (age <40 years; 18 subjects). Serum was analyzed with 2-dimensional electrophoresis and Western blot, with cultured human brain microvascular endothelial cells as the antigen source. The anti-Tom40 antibody was identified significantly more frequently in patients with Alzheimer's disease than control subjects or patients with multiple sclerosis. In patients with Alzheimer's disease, the mean scores for the Mini-Mental State Examination were significantly lower for patients who were positive for anti-Tom40 antibody than those who were negative for anti-Tom40 antibody. In summary, the anti-Tom40 antibody is significantly associated with cognitive impairment in patients with Alzheimer's disease.

The measurement of serum ceruloplasmin is useful for diagnostic differentiation of immune thrombocytopenic purpura.

BACKGROUND: To expand the criteria of immune thrombocytopenic purpura (ITP), which is a diagnosis of exclusion, we analyzed proteins separated by 1-dimensional gel electrophoresis of the retained fraction in a Con A-Sepharose column from ITP patients' sera. METHODS: Serum samples were from 19 adult patients with chronic ITP, 9 patients with thrombocytopenia of decreased production, and 20 healthy controls. Samples were applied to a Con A-Sepharose column, and the retained fraction was subjected to 10% SDS-PAGE. The % area of each densitometric protein peak was compared between the two groups and proteins in each band were identified using LC-MS/MS. RESULTS: Eleven protein bands were distinctly separated by 1-dimensional electrophoresis. The percent area of bands #2 and #3 were significantly higher in ITP patients than in controls. The percent area of band #2 (p<0.01) was significantly higher in ITP patients than in non-ITP patients. We identified alpha(2)-macroglobulin, ceruloplasmin (Cp), and C3 in band #2 and complement factor B in #3 band. Serum concentrations of alpha(2)-macroglobulin and Cp were significantly higher in ITP patients than in controls. Serum concentrations of Cp were significantly higher in ITP patients than in non-ITP patients (p=0.0005). Serum complement factor B concentrations were significantly higher in ITP patients and non-ITP patients than in controls. ROC analysis showed that the total percent area of bands #2 and #3, and Cp had higher diagnosis availability for ITP patients when compared with controls and non-ITP patients, respectively. CONCLUSIONS: Measurement of Cp separated by the present method could be useful for the diagnosis of ITP in the presence of thrombocytopenia and a non- or low-inflammatory state.