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Complement component 9 deficiency is not associated with the development of gastric carcinoma and leukaemia
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Abstracts / Molecular Immunology 45 (2008) 4095–4182
risk factor for disease susceptibility of SLE among the three races studied. doi:10.1016/j.molimm.2008.08.241 P243 Cytokine imbalance and cross-talks among cytokines, immune complexes and genetic factors on complement receptor 1 expression in health and disease (systemic lupus erythematosus) Nibhriti Das a , Vaishali Arora a , Vishal Marwah a , Jyotsna Verma b , Rahul Gover a , Ashok Kumar a , Devyani Anand a , Bintili Biswas a , Uma Kumar a a b
All India Institute of Medical Sciences, New Delhi, India Sir Ganga Ram Hospital, New Delhi, India
Decline in neutrophil CR1 transcript in SLE had been reported. To test an interlinked influence of cytokines, CR1 gene polymorphism and immune complexes on CR1 expression, we monitored the serum levels of IFN-␥, IL-4, TNF-␣, IL-10 & circulating Immune Complex (CIC), evaluated their correlations in 40 healthy individuals and 40 patients with SLE, and studied the effect of these factors on leukocyte CR1 expression by in vitro culture studies. The levels of TNF-␣ and IL-10 were higher (P < 0.001, 6.9 times & 2.9 times mean values 632.72 ± 118.36 pg/ml/91.43 ± 30.91 & 196.81 ± 75.84 pg/ml/166.24 ± 29.75 pg/ml in patients/normals, respectively, alteration in the levels of the rest were not significant. A positive correlation (r = 0.327, p < 0.05) was observed between the levels of IL-10 and TNF-␣ in patients but not in normals. The correlation between IL-10 vs. IL-4 (r = 0.345) was positive; and between IL-10 vs. IFN-␥ (r = −0.382) was negative and significant (p < 0.05) in patients but not in normals. Levels of IL-4 and IFN-␥ gave a highly significant (p < 0.001) positive correlation both in normals (r = −0.987) as well as patients (r = −0.990). A negative correlation (r = −0.363, p < 0.05) was observed between IL-10 and leucocyte CR1 (L-CR1) transcript only in patients. In vitro studies revealed that of all the cytokines, IFN-␥ was the only one to significantly increase the levels of NCR1 transcript in patients and controls. Heat aggregated IgG (Hagg.IgG) reduced the CR1 transcript levels. IL-4, IL-10 and Hagg.IgG antagonized the effect of IFN-␥. TNF-␣ had no effect on CR1 expression. The effects of IFN-␥ and serum opsonized Hagg.IgG on CR1 expression was more pronounced in patients and controls of HL and LL genotypes. The observations suggested SLE. IL-10 appeared as the key player in the scenario. Findings suggest functional imbalance in the cytokine network and integrated effect of cytokines, immune complex and CR1 genotypes in modulating the levels of CR1 in SLE. doi:10.1016/j.molimm.2008.08.242 P244 Complement component 9 deficiency is not associated with the development of gastric carcinoma and leukaemia Rie Kuroki-Nagamatsu, Yuhki Koga, Aiko Suminoe, Akinobu Matsuzaki, Kenji Ihara, Eiji Oki, Yoshihiro Kakeji, Yoshihiko Maehara, Toshiro Hara Kyushu University, Fukuoka, Japan Patients with certain primary immunodeficiency diseases have a predisposition to the development of cancer, whereas it is not known whether congenital complement deficiency is a risk factor for the development of malignancies. Deficiency of the ninth
component of complement (C9) is the most common complement deficiency in Japanese with an incidence of approximately one homozygote in 1000 individuals (0.1%). We previously reported that C9 deficiency showed an increased risk for meningococcal meningitis [J. Pediatr., 1989], but not for SLE [Lupus, 2000], and that almost all C9 deficiency in Japanese was caused by a common mutation of C-to-T transition leading to TGA stop codon for Arg95 (R95X) in exon 4 of the C9 gene [Hum. Genet., 1998]. The aim of this study was to assess any contribution of C9 deficiency to the development of 2 representative epithelial and non-epithelial malignancies, gastric carcinoma and lymphoid malignancy, which occur in association with primary immunodeficiencies in Japan. The genotyping was carried out by the allele-specific polymerase chain reaction (ASPCR) method, which was established in our previous paper [J. Hum. Genet., 1999]. As a result, the heterozygous genotype of R95X was identified in 4 of 91 gastric carcinoma patients (4.4%) and in 3 of 64 childhood leukemia patients (4.7%). No homozygotes for the R95X were found in the cancer populations. The R95X carrier frequencies showed no significant differences between patients with gastric carcinoma or leukaemia and controls (6.7%), strongly suggesting that the frequency of C9 deficiency in patients with gastric carcinoma or leukaemia was similar to that in controls. Considering that a high frequency of C9 deficiency is found among patients with meningococcal meningitis, we conclude that C9 deficiency does not make a great contribution to the development of gastric carcinoma and leukaemia in Japanese. doi:10.1016/j.molimm.2008.08.243 P245 Complement components as biomarkers of disease in multiple sclerosis Gillian Ingram, Svetlana Hakobyan, Claire Hirst, Neil Robertson, B. Paul Morgan School of Medicine, Cardiff University, Cardiff, UK Disease course in multiple sclerosis (MS) can be difficult to define with available clinical tools but is of importance in guiding interventions. Effective and accessible biomarkers are required. We examined whether measurement of complement components distinguished MS disease subgroups. C3, C4, factor B (fB), and regulator factor H (fH) along with its Y402H polymorphic variants (implicated in other chronic inflammatory CNS conditions) were measured in a large, well-characterised cohort of MS patients and matched controls. C3 and C4 were measured by nephelometry while fB, fH and fH variants were measured by ELISA using newly developed monoclonal antibodies with purified proteins as standards. C3, C4 and fB were measured in 80 patients, while fH and variants were measured in 258 patients, all classified according to disease course with subsequent samples in individuals over 2 years. After controlling for disease duration, treatment, age and disability, fH levels were upregulated in progressive disease (p < 0.001) and down-regulated in relapsing disease (p = 0.001) compared to controls. In sequentially sampled patients, acute relapse resulted in transiently increased fH levels (p < 0.001). In clinically stable patients sampled sequentially, fH levels remained constant over one year (CV% = 6.8). Although there was no difference in the Y402H allele frequency in MS patients compared to controls (p = 0.56), the concentration of the H402 variant was lower in stable relapsing disease (p = 0.002). Y402H genetic status had no effect on fH levels. FB levels were negatively correlated with fH levels (r2 = 0.082/p = 0.002). There were no significant changes in C3 and C4 levels between MS subgroups or with con-
Abstracts / Molecular Immunology 45 (2008) 4095–4182
risk factor for disease susceptibility of SLE among the three races studied. doi:10.1016/j.molimm.2008.08.241 P243 Cytokine imbalance and cross-talks among cytokines, immune complexes and genetic factors on complement receptor 1 expression in health and disease (systemic lupus erythematosus) Nibhriti Das a , Vaishali Arora a , Vishal Marwah a , Jyotsna Verma b , Rahul Gover a , Ashok Kumar a , Devyani Anand a , Bintili Biswas a , Uma Kumar a a b
All India Institute of Medical Sciences, New Delhi, India Sir Ganga Ram Hospital, New Delhi, India
Decline in neutrophil CR1 transcript in SLE had been reported. To test an interlinked influence of cytokines, CR1 gene polymorphism and immune complexes on CR1 expression, we monitored the serum levels of IFN-␥, IL-4, TNF-␣, IL-10 & circulating Immune Complex (CIC), evaluated their correlations in 40 healthy individuals and 40 patients with SLE, and studied the effect of these factors on leukocyte CR1 expression by in vitro culture studies. The levels of TNF-␣ and IL-10 were higher (P < 0.001, 6.9 times & 2.9 times mean values 632.72 ± 118.36 pg/ml/91.43 ± 30.91 & 196.81 ± 75.84 pg/ml/166.24 ± 29.75 pg/ml in patients/normals, respectively, alteration in the levels of the rest were not significant. A positive correlation (r = 0.327, p < 0.05) was observed between the levels of IL-10 and TNF-␣ in patients but not in normals. The correlation between IL-10 vs. IL-4 (r = 0.345) was positive; and between IL-10 vs. IFN-␥ (r = −0.382) was negative and significant (p < 0.05) in patients but not in normals. Levels of IL-4 and IFN-␥ gave a highly significant (p < 0.001) positive correlation both in normals (r = −0.987) as well as patients (r = −0.990). A negative correlation (r = −0.363, p < 0.05) was observed between IL-10 and leucocyte CR1 (L-CR1) transcript only in patients. In vitro studies revealed that of all the cytokines, IFN-␥ was the only one to significantly increase the levels of NCR1 transcript in patients and controls. Heat aggregated IgG (Hagg.IgG) reduced the CR1 transcript levels. IL-4, IL-10 and Hagg.IgG antagonized the effect of IFN-␥. TNF-␣ had no effect on CR1 expression. The effects of IFN-␥ and serum opsonized Hagg.IgG on CR1 expression was more pronounced in patients and controls of HL and LL genotypes. The observations suggested SLE. IL-10 appeared as the key player in the scenario. Findings suggest functional imbalance in the cytokine network and integrated effect of cytokines, immune complex and CR1 genotypes in modulating the levels of CR1 in SLE. doi:10.1016/j.molimm.2008.08.242 P244 Complement component 9 deficiency is not associated with the development of gastric carcinoma and leukaemia Rie Kuroki-Nagamatsu, Yuhki Koga, Aiko Suminoe, Akinobu Matsuzaki, Kenji Ihara, Eiji Oki, Yoshihiro Kakeji, Yoshihiko Maehara, Toshiro Hara Kyushu University, Fukuoka, Japan Patients with certain primary immunodeficiency diseases have a predisposition to the development of cancer, whereas it is not known whether congenital complement deficiency is a risk factor for the development of malignancies. Deficiency of the ninth
component of complement (C9) is the most common complement deficiency in Japanese with an incidence of approximately one homozygote in 1000 individuals (0.1%). We previously reported that C9 deficiency showed an increased risk for meningococcal meningitis [J. Pediatr., 1989], but not for SLE [Lupus, 2000], and that almost all C9 deficiency in Japanese was caused by a common mutation of C-to-T transition leading to TGA stop codon for Arg95 (R95X) in exon 4 of the C9 gene [Hum. Genet., 1998]. The aim of this study was to assess any contribution of C9 deficiency to the development of 2 representative epithelial and non-epithelial malignancies, gastric carcinoma and lymphoid malignancy, which occur in association with primary immunodeficiencies in Japan. The genotyping was carried out by the allele-specific polymerase chain reaction (ASPCR) method, which was established in our previous paper [J. Hum. Genet., 1999]. As a result, the heterozygous genotype of R95X was identified in 4 of 91 gastric carcinoma patients (4.4%) and in 3 of 64 childhood leukemia patients (4.7%). No homozygotes for the R95X were found in the cancer populations. The R95X carrier frequencies showed no significant differences between patients with gastric carcinoma or leukaemia and controls (6.7%), strongly suggesting that the frequency of C9 deficiency in patients with gastric carcinoma or leukaemia was similar to that in controls. Considering that a high frequency of C9 deficiency is found among patients with meningococcal meningitis, we conclude that C9 deficiency does not make a great contribution to the development of gastric carcinoma and leukaemia in Japanese. doi:10.1016/j.molimm.2008.08.243 P245 Complement components as biomarkers of disease in multiple sclerosis Gillian Ingram, Svetlana Hakobyan, Claire Hirst, Neil Robertson, B. Paul Morgan School of Medicine, Cardiff University, Cardiff, UK Disease course in multiple sclerosis (MS) can be difficult to define with available clinical tools but is of importance in guiding interventions. Effective and accessible biomarkers are required. We examined whether measurement of complement components distinguished MS disease subgroups. C3, C4, factor B (fB), and regulator factor H (fH) along with its Y402H polymorphic variants (implicated in other chronic inflammatory CNS conditions) were measured in a large, well-characterised cohort of MS patients and matched controls. C3 and C4 were measured by nephelometry while fB, fH and fH variants were measured by ELISA using newly developed monoclonal antibodies with purified proteins as standards. C3, C4 and fB were measured in 80 patients, while fH and variants were measured in 258 patients, all classified according to disease course with subsequent samples in individuals over 2 years. After controlling for disease duration, treatment, age and disability, fH levels were upregulated in progressive disease (p < 0.001) and down-regulated in relapsing disease (p = 0.001) compared to controls. In sequentially sampled patients, acute relapse resulted in transiently increased fH levels (p < 0.001). In clinically stable patients sampled sequentially, fH levels remained constant over one year (CV% = 6.8). Although there was no difference in the Y402H allele frequency in MS patients compared to controls (p = 0.56), the concentration of the H402 variant was lower in stable relapsing disease (p = 0.002). Y402H genetic status had no effect on fH levels. FB levels were negatively correlated with fH levels (r2 = 0.082/p = 0.002). There were no significant changes in C3 and C4 levels between MS subgroups or with con-