Complement C3 glomerulopathy refers to a disease process in which abnormal control of Complement activation or degradation results in predominant C3 fragment deposition within the glomerulus and causes glomerular damage. Abnormal control of the Complement alternative pathway is a well-established risk factor for the occurrence of C3 glomerulonephritis. It is the first reported case in Iran with multiple mutations in Complement factor H, with one of these mutations we have expected in hemolytic uremic syndrome rather than C3 glomerulopathy. Genetic analysis showed that the molecular abnormalities of factor H led to Complement factor H malfunction that were polymorphous and not restricted to the C-terminal domains of the protein.

While terminal Complement blockade with Eculizumab is the first line therapy for atypical HUS (Haemolytic uremic syndrome), lack of its availability and cost limits its use. Plasma exchange becomes a first line modality in the current scenario. However, exposure to large volumes of allogenic plasma and lack of skilled manpower are the limiting factors associated with it. Moreover, there is a subset of patients who fail to respond to plasma exchange. Treatment and follow up records of the children with atypical HUS who did not respond to daily plasma exchange therapy and their course during follow up was reviewed. Three children with positive anti-Complement factor H antibody atypical HUS did not respond to daily plasma exchange and were administered Rituximab after completing five daily plasma exchange therapies. It was seen that these children, initially non-responsive to plasma therapy, attained remission after Rituximab and did not require further plasma exchanges. The remission was sustained in long term with a follow up of 7 years, 4 years and 6 months respectively. Rituximab might be a useful alternative in inducing haematological remission in children with poor or no response to plasma therapy. This abbreviates the duration of plasma exchange, which not only avoids complications due to prolonged plasma therapy but also helps reducing the cost of therapy.

Background: Neovascular age-related macular degeneration (nAMD) is one of the main causes of blindness in developed countries. Complement factor H (CFH) is one of the genes involved in the pathogenesis of nAMD. This study investigated the rs10737680 polymorphism in CFH and its conferred susceptibility to nAMD in Yogyakarta, Indonesia. Methods: This case-control hospital-based study recruited participants consisting of 96 patients with nAMD and 101 controls without nAMD from the Eye Polyclinic of Sardjito Hospital, YAP Eye Hospital, and Hardjolukito Hospital Yogyakarta. nAMD was diagnosed when fundus examination, fundus photographs, and optical coherence tomography revealed hard or soft drusen in the macular area measuring > 63 µ, m that appeared below the retinal pigment epithelium, with or without macular hypo-or hyperpigmentation, and was accompanied by choroidal neovascularization. Genomic DNA was extracted using a commercial DNA isolation kit. The restriction fragment length polymorphism technique was used to identify the rs10737680 polymorphism in CFH. Results: The mean (standard deviation [SD]) age of the nAMD group was not homogeneous with that of the control group (P < 0. 05),65. 41 (9. 74) years versus 68. 24 (7. 82) years. The number of patients with hypertension in the nAMD group was significantly higher than in the control group (P < 0. 05). In the nAMD group, the genotype distribution indicated homozygous risk allele in 34. 38%, heterozygous risk allele in 57. 29%, and homozygous non-risk allele in 8. 33%. In the control group, the genotype distribution indicated homozygous risk allele in 21. 78%, heterozygous risk allele in 36. 63%, and homozygous non-risk allele in 41. 58%. Statistical analysis between the two study groups according to homozygous risk allele genotype (odds ratio [OR], 7. 87,95% confidence interval [CI], 2. 88–, 22. 79) and heterozygous genotype (OR, 7. 80,95% CI, 3. 11–, 21. 19) showed a significant difference (both P < 0. 01). Conclusions: Homozygous risk allele was less frequent than heterogeneous risk allele in patients with nAMD,however, both increased the risk for nAMD. Although the homozygous or heterozygous risk-alleles were detected in most patients, yet other important genetic or environmental factors could be involved in the pathogenesis of nAMD. Overall, we found a significant association between rs10737680 polymorphism in CFH and the susceptibility to nAMD in Yogyakarta, Indonesia,however, future studies are needed to fully delineate the mechanism.

Membranoproliferative glomerulonephritis (MPGN) is characterized by proliferation of mesangial and endothelial cells and by thickening of the peripheral capillary walls. Type II of the MPGN is associated with Complement abnormalities which are factor H deficiencies due to mutations in the Complement factor H (CFH) gene. We report a 15-year-old boy diagnosed with MPGN II in whom genetic analyses of the CFH gene revealed that the patient was heterozygote for a polymorphism in exon 2 of the CFH (c.184G>A), heterozygote for a polymorphism in exon 9 of the CFH (c.1204C>T), and heterozygote for a polymorphism in exon 10 of the CFH (c.1419G>A). These data recapitulate a prototypical Complement genetic profile, the presence of major risk factors for MPGN II, which support the hypothesis that these dense deposit diseases have a common pathogenic mechanism involving dysregulation of the alternative pathway of Complement activation.

Introduction. Thrombotic microangiopathy (TMA) is a unifying term for both hemolytic uremic syndrome (HUS) and thrombotic thrombocytopenia purpura (TTP). TMA is characterized by thrombocytopenia, hemolytic anemia, and microvascular thrombi in different organ systems. Known risk factors for development of TMA including bacterial toxins, viral infection, autoimmune disease, and pregnancy catastrophes. Inherited deficiency of Complement inhibitory factor H (FH) and metalloproteinase ADAMTS-13 (vWF-cleaving protease) deficiencies or inhibitors are another important risk factor for development of TMA and may coexist with above risk factors. Methods. In this single center prospective study, all patients with clinical diagnosis of TMA were entered. Patients’ clinical presentation and routine laboratory measurements were re-ordered and serum samples were taken for measurement of Complement inhibitory factor H, ADAMTS-13 activity, and presence or absence of ADAMTS-13 inhibitor. Serum Complement C3, C4, CH50 level, serologic test for hepatitis B, hepatitis C, HIV, systemic lupus erythematous, and rheumatoid arthritis were performed. History of recent diarrhea and obstetric complications was recorded. We followed recurrence of TMA and renal outcome of these patients for two years.Results. Between 2003 and 2005, we diagnosed 11 consecutive TMA patients (f/m 10/1, age; 22 to 70 years). Majority of our patients were young women. Routine laboratory measurements were as the following: hemoglobin: 8.8 ± 0.9 mg/dl, platelet count: 40800 ± 14793/ml, LDH: 1793 ± 585 IU/L, C3: 195 ± 60mg/ dl, C4: 51 ± 11mg/dl, Complement factor H: 772 ± 211 mg/dl, and ADAMTS-13 activity percentage: 32.2% ± 13.7% (range, 50% to 150% activity). One patient had ADAMTS-13 inhibitory factor. Acute dialysis was started in four patients (37%). One patient had a positive history of diarrhea. Two patients (2/11) had systemic lupus erythematous. HBS-Ag and HCV antibody positive results were detected in two patients. Obstetric catastrophes, including IUFD and sever preeclampsia were detected in two patients. One patient with ADAMTS-13 inhibitory factor who presented with unconsciousness and convulsion died in his first week of admission. Recurrent TMA occurred in a patient with low C3 Complement level. Chronic Renal dysfunction was developed in two patients. Conclusion. In our study, majority of the patients with TMA were young women. Patients with ADAMTS-13 inhibitory factors and Complement factors deficiency had a poor prognosis outcome.

Background: Age-related macular degeneration (AMD) is the leading cause of vision loss in the elderly. Although it has been shown that Y402H polymorphism in the CFH gene was strongly associated with AMD in the Iranian population, there were no data on other single nucleotide polymorphisms (SNPs), which have the most significant association with AMD. This study aimed to investigate hot point regions in exon 10 and intron 9. Materials and Methods: One hundred and sixty-six AMD patients and 69 controls were recruited. Their blood was collected in the tubes containing EDTA. Then, DNA was extracted from the blood, and its quality was evaluated. Primers were designed for intron 9 and exon10 sequencing. A viral polymorphisms analysis software named CEQ was used for the analysis of putative polymorphisms. Results: We noticed three polymorphisms in study cases: rs7535263 and C66379A in intron 9 and rs2274700 in exon 10. Based on the McNamara’ s test (rs7535263 and rs2274700) and the Phi and Cramer’ s test (C66379A), a significant difference was found between the control and patient groups regarding rs7535263 and rs2274700 polymorphisms. Conclusion: We found a synonymous or silent mutation, A473A, rs2274700 in exon 10 in 85% of patients. From two intronic SNPs, just rs7535263 showed association with the disease in studied patients living in Gilan Province, Iran. Although no significant relationship was found between controls and patients regarding the C66379A allele, it would be important that no other sources have reported C66379A polymorphism in AMD yet.

Background and Objective: Neisseria meningitidis is a leading cause of meningitis and sepsis worldwide. The factor H binding protein (fHBP) is a key virulence factor of Neisseria meningitidis that is able to selectively bind to human factor H, the key regulator of the alternative Complement pathway, which it has important implications for meningococcal pathogenesis and vaccine design. The aims of present research were cloning, expression, purification of fHbp and confirmation of the interaction between serum factor H (fH) and produced factor H binding protein. Materials and Methods: A 820 base pairs fhbp gene fragment was amplified by PCR and cloned into expression vector pET28a (+) in Bam HI and SalI restriction enzymes sites. Recombinant DNA was expressed in BL21 (DE3) cell. fHBP protein was purified by Ni-NTA agarose resin. Coupling of recombinant protein into CNBr activated Sepharose 4B resin was carried out for application in serum fH protein purification. (fH-fHBP) interaction was confirmed by SDS-PAGE and far-western blotting.Results and Conclusions: SDS-PAGE results showed a 35 kDa protein band. 150 kDa fH protein was purified by designed Sepharose 4B resin. Far-western blotting confirmed (fH-fHBP) interaction and proper folding of factor H binding protein.