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Reference level for factor H autoantibodies in the Japanese population using a standardized enzyme-linked immunosorbent assay
Abstracts / Immunobiology 221 (2016) 1131–1225
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Reference level for factor H autoantibodies in the Japanese population using a standardized enzyme-linked immunosorbent assay
The analyzation of complement deposition in tauopathies
Yusuke Okuda 1,∗ , Toshiki Masuda 1 , Tomoyuki Sakai 1 , Toshinaga Maeda 2 , Toshihiro Sawai 1 1 Department of Pediatrics, Shiga University of Medical Science, Shiga, Japan 2 Division of Molecular Medical Biochemistry, Department of Biochemistry and Molecular Biology, Shiga University of Medical Science, Shiga, Japan
Background: Detecting factor H autoantibodies (FH Abs) is essential for diagnosis and therapy of atypical hemolytic uremic syndrome (aHUS). A standardized enzyme-linked immunosorbent assay (ELISA) for FH Abs was recently established by the European working group. To apply the standardized method for clinical practice in Japan, we attempted to determine the Japanese reference level for FH Abs. Methods: We recruited healthy volunteers aged ≥20 years. “Healthy” was defined as no past history of kidney disease, including macrohematuria, or purpura. The FH Ab-positive sample used for the standard curve was plasma from a patient with aHUS who had an FH Ab titer of 16,000 arbitrary units (AU)/mL. The standard curve was generated from samples diluted from 1/1600 to 1/204,800. The positive control sample was serum from a patient with dense deposit disease and an FH Ab titer of 92 AU/mL. Sera from the participants were diluted 1/50 and we set the normal range as the mean ± 2 standard deviations. ELISA was performed according to a previously published standardized method. C3, C4 and CH50 levels of simultaneously obtained sera were also measured. Results: Fifty volunteers (22 males, 28 females) were recruited. The median age was 31.5 years old (range 22–52). Mean C3, C4 and CH50 levels were 94.1 ± 19.0, 21.2 ± 5.9 and 37.5 ± 7.5 mg/dL, respectively. The mean optical density (OD) was 0.22 ± 0.11 (standard sample level: 1.94 for 1/1,600 to 0.06 for 1/204,800). The mean FH Ab level in the healthy participants was 14.1 ± 8.1 AU/mL. Conclusion: The upper limit titer of FH Abs in the Japanese population by standardized ELISA was 30 AU/mL, which is different from previously reported results. Whether this difference is only due to the standard sample used or is due to racial difference is unclear. To clarify this issue, we are planning future studies to investigate sample types used frequently as standards in previous studies. http://dx.doi.org/10.1016/j.imbio.2016.06.147
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Hiroyasu Akatsu 1,2,3,∗ , Norihiro Ogawa 3 , Takeshi Kanesaka 3 , Hirotaka Ohara 1 , Noriyuki Matsukawa 2 , Hideaki Suzuki 4 , Takashi Asasa 5 , Kazuhiko Uchida 4,6 , Yoshio Hashizume 3 1 Department of Community-Based Medical Education, Nagoya City University Graduate School of Medical Sciences, Nagoya, Aichi, Japan 2 Department of Neurology and Neuroscience, Nagoya City University Graduate School of Medical Sciences, Nagoya, Aichi, Japan 3 Department of Neuropathology, Choju Medical Institute, Fukushimura Hospital, Toyohashi, Aichi, Japan 4 MCBI (Molecular and Clinical Bioinformatics), Tsukuba, Ibaraki, Japan 5 Tokyo Medical and Dental University, Faculty of Medicine, Tokyo, Japan 6 Faculty of Medicine, University of Tsukuba, Ibaraki, Japan
Introduction: One of the most common dementia is Alzheimer’s disease (AD), also as neurodegenerative disease. Its main neuropathological findings are senile plaques (SPs) and neurofibrillary tangles (NFTs). There are several dementias as neurodegenerative disease with phosphorylated tau (p-tau) which is one of the components of NFTs. In AD brain, the complement activation is observed mainly in SPs but also in NFTs. But there are a few report focused in p-tau with AD and another neurodegenerative disease (tauopathies). In this report, we analyze complement deposition in human brain with non-AD tauopathies, for example tangle only dementia (TD), argyrophilic grain disease (AGD), progressive supranuclear palsy (PSP), and corticobasal degeneration (CBD). Materials and methods: The human brain samples are autopsy confirmed human brains stocked Fukushimura Brain Bank approved by the Ethics Committee. The immuno-histochemical analysis were used with antibodies of anti-human C1q, C3 and C4d to detect complement deposition in buffered 10% formaldehyde fixed brain which embedded paraffin wax. Results: In TD’s brain around NFTs, the C1q, C3 and C4d are weakly deposited like AD. Need analyzation of quantification in future, the complement activation in NFTs in TD are stronger than AD. In another tauopaties, although we have to check more details but it seems that few complement responsibilities occurred. Conclusions: The complement activations in NFTs are weaker than in SPs but in NFTs of TDs without SPs, the complement factors are more deposited in AD brain. In these results, NFTs of TDs could strongly have complement response than another tauopathies. http://dx.doi.org/10.1016/j.imbio.2016.06.148
132
133
Reference level for factor H autoantibodies in the Japanese population using a standardized enzyme-linked immunosorbent assay
The analyzation of complement deposition in tauopathies
Yusuke Okuda 1,∗ , Toshiki Masuda 1 , Tomoyuki Sakai 1 , Toshinaga Maeda 2 , Toshihiro Sawai 1 1 Department of Pediatrics, Shiga University of Medical Science, Shiga, Japan 2 Division of Molecular Medical Biochemistry, Department of Biochemistry and Molecular Biology, Shiga University of Medical Science, Shiga, Japan
Background: Detecting factor H autoantibodies (FH Abs) is essential for diagnosis and therapy of atypical hemolytic uremic syndrome (aHUS). A standardized enzyme-linked immunosorbent assay (ELISA) for FH Abs was recently established by the European working group. To apply the standardized method for clinical practice in Japan, we attempted to determine the Japanese reference level for FH Abs. Methods: We recruited healthy volunteers aged ≥20 years. “Healthy” was defined as no past history of kidney disease, including macrohematuria, or purpura. The FH Ab-positive sample used for the standard curve was plasma from a patient with aHUS who had an FH Ab titer of 16,000 arbitrary units (AU)/mL. The standard curve was generated from samples diluted from 1/1600 to 1/204,800. The positive control sample was serum from a patient with dense deposit disease and an FH Ab titer of 92 AU/mL. Sera from the participants were diluted 1/50 and we set the normal range as the mean ± 2 standard deviations. ELISA was performed according to a previously published standardized method. C3, C4 and CH50 levels of simultaneously obtained sera were also measured. Results: Fifty volunteers (22 males, 28 females) were recruited. The median age was 31.5 years old (range 22–52). Mean C3, C4 and CH50 levels were 94.1 ± 19.0, 21.2 ± 5.9 and 37.5 ± 7.5 mg/dL, respectively. The mean optical density (OD) was 0.22 ± 0.11 (standard sample level: 1.94 for 1/1,600 to 0.06 for 1/204,800). The mean FH Ab level in the healthy participants was 14.1 ± 8.1 AU/mL. Conclusion: The upper limit titer of FH Abs in the Japanese population by standardized ELISA was 30 AU/mL, which is different from previously reported results. Whether this difference is only due to the standard sample used or is due to racial difference is unclear. To clarify this issue, we are planning future studies to investigate sample types used frequently as standards in previous studies. http://dx.doi.org/10.1016/j.imbio.2016.06.147
1191
Hiroyasu Akatsu 1,2,3,∗ , Norihiro Ogawa 3 , Takeshi Kanesaka 3 , Hirotaka Ohara 1 , Noriyuki Matsukawa 2 , Hideaki Suzuki 4 , Takashi Asasa 5 , Kazuhiko Uchida 4,6 , Yoshio Hashizume 3 1 Department of Community-Based Medical Education, Nagoya City University Graduate School of Medical Sciences, Nagoya, Aichi, Japan 2 Department of Neurology and Neuroscience, Nagoya City University Graduate School of Medical Sciences, Nagoya, Aichi, Japan 3 Department of Neuropathology, Choju Medical Institute, Fukushimura Hospital, Toyohashi, Aichi, Japan 4 MCBI (Molecular and Clinical Bioinformatics), Tsukuba, Ibaraki, Japan 5 Tokyo Medical and Dental University, Faculty of Medicine, Tokyo, Japan 6 Faculty of Medicine, University of Tsukuba, Ibaraki, Japan
Introduction: One of the most common dementia is Alzheimer’s disease (AD), also as neurodegenerative disease. Its main neuropathological findings are senile plaques (SPs) and neurofibrillary tangles (NFTs). There are several dementias as neurodegenerative disease with phosphorylated tau (p-tau) which is one of the components of NFTs. In AD brain, the complement activation is observed mainly in SPs but also in NFTs. But there are a few report focused in p-tau with AD and another neurodegenerative disease (tauopathies). In this report, we analyze complement deposition in human brain with non-AD tauopathies, for example tangle only dementia (TD), argyrophilic grain disease (AGD), progressive supranuclear palsy (PSP), and corticobasal degeneration (CBD). Materials and methods: The human brain samples are autopsy confirmed human brains stocked Fukushimura Brain Bank approved by the Ethics Committee. The immuno-histochemical analysis were used with antibodies of anti-human C1q, C3 and C4d to detect complement deposition in buffered 10% formaldehyde fixed brain which embedded paraffin wax. Results: In TD’s brain around NFTs, the C1q, C3 and C4d are weakly deposited like AD. Need analyzation of quantification in future, the complement activation in NFTs in TD are stronger than AD. In another tauopaties, although we have to check more details but it seems that few complement responsibilities occurred. Conclusions: The complement activations in NFTs are weaker than in SPs but in NFTs of TDs without SPs, the complement factors are more deposited in AD brain. In these results, NFTs of TDs could strongly have complement response than another tauopathies. http://dx.doi.org/10.1016/j.imbio.2016.06.148