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Systemic lupus erythematosus (SLE) is a systemic autoimmune disorder often occurring in women of childbearing age. SLE is characterized by pathogenic antibody production and inflammation. Histone deacetylase (HDAC) 6 is a class IIb histone deacetylase member. HDAC6 has the ability to catalyze the removal of acetyl groups from lysine residues on non-histone proteins. It has been observed that in lupus mouse models, specific HDAC6 inhibition reduces inflammation. Administration of pristane, a naturally occurring hydrocarbon oil, can result in lupus-like illness and persistent inflammation. In our studies, 0.5 ml of pristane or phosphate buffered saline (PBS) was given intraperitoneally into sex- and age-matched wild type (WT) and HDAC6-/- mice on the C57BL/6 background at 8–12 weeks of age, and mice were euthanized 10 days or 8 months later. The animals were assessed as they aged. Short-term pristane treatment promoted the population of CD11b+Ly6C++ inflammatory monocytes and CD11b+Ly6G+ neutrophils. Peritoneal recruitment of these inflammatory monocytes and neutrophils in HDAC6-/- mice was significantly decreased compared to the WT mice. Pristane treatment also induced the interferon (IFN) signature genes as determined by RT-qPCR. Furthermore, IFN signature genes were decreased in HDAC6-/- mice compared to the WT mice. In vitro studies in J774 cells revealed that the selective HDAC6 inhibitor (ACY-738) increased acetylation of NF-κB while increasing STAT1-phosphorylation which caused the synthesis of inducible nitric oxide synthase (iNOS) in cells activated by LPS and IFN-γ. Long-term pristane treatment induced proteinuria in female mice although there were no significant differences between WT and HDAC6-/- animals. HDAC6 deletion significantly inhibited anti-double stranded (ds) DNA IgG level compared with WT mice. Moreover, HDAC6 deletion decreased some lymphocyte populations like T-helper 17 (Th17) cells after pristane treatment while not affecting other cell populations, such as regulatory T cells, total T cells, B cells, and plasma cells. Taken together, these results demonstrate that although HDAC6 inhibition may inhibit some inflammatory pathways, others remain unaffected. / Doctor of Philosophy / Systemic lupus erythematosus (SLE) is an autoimmune disorder that affects the entire body. It's more common in women of childbearing age. SLE involves the immune system attacking healthy tissues leading to inflammation. One hallmark is the production of autoantibodies. SLE can affect various organs and tissues, causing symptoms like joint pain, skin rashes, and fatigue. Histone Deacetylase 6 (HDAC6) is a specific protein involved in modifying protein function by removing acetyl groups. In lupus, inhibiting HDAC6 has been reported to reduce inflammation. Pristane, a natural oil, can trigger lupus-like symptoms and persistent inflammation. In our studies, we investigated the role of HDAC6 on pristane induced lupus. We used both normal mice (WT) and mice lacking HDAC6 (HDAC6-/-). Mice were injected with pristane or a control solution. After 10 days or 8 months, we assessed the mice. We found 10-day pristane treatment increased inflammatory monocytes and neutrophils. HDAC6-/- mice had fewer of these immune cells in their peritoneum. Pristane also activated interferon genes, but this effect was reduced in HDAC6-/- mice. In our studies, a HDAC6 inhibitor increased the acetylation of NF-κB (that would dampen inflammation). Eight-month pristane administration induced proteinuria (protein in urine) in female mice, and this is true for both WT and HDAC6-/- mice. However, HDAC6 deletion decreased autoantibody levels and a pro-inflammatory cell type called Th17. In conclusion, HDAC6 plays a role in lupus-related inflammation. Targeting HDAC6 might be a potential therapeutic approach for managing lupus symptoms.

Orientador : José Gastao Rocha de Carvalho / Co-orientador : Valderílio Feijó Azevedo / Dissertaçao (mestrado) - Universidade Federal do Paraná, Setor de Ciencias da Saúde, Programa de Pós-Graduação em Medicina Interna / Resumo: A fim de se caracterizar a nefrite lúpica incipiente, foram analisadas suas manifestações clínicas, dosada a microalbuminúria (jjALB) e realizada biópsia renal em 30 pacientes portadores de lúpus eritematoso sistêmico (LES) sem evidência clínico-laboratorial de comprometimento renal. As biópsias renais foram classificadas de acordo com a classes propostas pela OMS. Quinze pacientes (50%) apresentaram glomerulonefrite mesangial (GNM) classe llb; 12 apresentaram GNM classe lia e 3 não apresentaram alteração na microscopia óptica (MO) ou na imunofluorescência (IF) (classe I). Nenhum paciente apresentou alteração compatível com glomerulonefrite proliferativa ou membranosa. Depósitos fluorescentes anti-lgM foram observados em 83% das biópsias renais, associados com depósitos menos intensos de IgG, IgA e C3. Pacientes com GNM classe Ha tinham média de idade de 36,3 anos e duração da doença de 3,25 anos; sua média da juALB foi de 9,35^/g/min. Pacientes com GNM classe llb tinham menor média de idade quando comparados aos da classe tia (26,04 anos vs. 36,3 anos) (p<0,029) e duração da doença de 4,8 anos; sua média da jyALB foi de 39,94/vg/min. Seis desses pacientes (6/15, 40 %) apresentaram anticorpos anti-dsDNA positivos, diferentemente da classe lia onde não houve positividade para anti-dsDNA (p<0,002, two tailed). O grupo com fjh lB anormal apresentou menor média de idade (p<0,029) e menores níveis de C3 (p<0,0098) quando comparados ao grupo com j/ALB normal. Esses resultados sugerem a prevalência elevada de GNM classe llb e de depósitos de IgM à imunofluorescência apesar do pequeno número de manifestações clínico-laboratoriais nestes pacientes, associação entre GNM classe llb e anticorpos anti-dsDNA, relação entre ^ALB anormal e diminuição dos níveis de C3 e que o nível de (jALB não discrimina a presença ou não de lesão renal na MO ou na IF nesta população. / Abstract: In an attempt to evaluate subclinical lupus nephropathy, we analysed the clinical characteristics, determined the albumin excretion rate (AER) by radioimmunoassay and performed renal biopsy in 30 patients with systemic lupus erythematosus (SLE) who had no clinical signs of renal involvement (no urinary sediment abnormalities, absence of proteinuria, serum creatinine <1.3 mg/dl). All biopsies were classified according to a modified classification proposed by the WHO. Fifteen cases (50%) had mesangial glomerulonephritis (MGN) type lib, 12 had MGN type lla and 3 patients showed no changes on light microscopy (LM) or on immunofluorescence (IF) (type I). None of the 30 patients had changes of membranous or diffuse proliferative glomerulonephritis. Anti-IgM fluorescent deposits were found in 83% of the renal biopsies, being associated with less heavily stained deposits of IgG, IgA and C3. Patients with MGN type lib showed lower mean-age when compared to those of MGN type lla (26,04 years vs. 36,3 years) (p<0,029); those patients also presented disease duration of 4.8 years and the mean AER was 39,94/jg/min. Six of the patients (6 of 15, 40%) showed positive anti-dsDNA antibodies, differently from patients with MGN type lla who didn't show positive anti-dsDNA antibodies (p<0,002, two-tailed). The group with abnormal AER presented lower mean-age (p<0,029) and lower C3 levels (p<0,0098) when compared to the group with normal AER. The results suggest the high prevalence of MGN type lib and IgM deposits on IF despite the paucity of clinical and iaboratorial data in these patients, the association between MGN type lib and positive anti-dsDNA antibodies, the relationship between abnormal AER and low C3 levels and the level of AER not being able to determine the presence or absence of renal disease on LM or IF in this population.

Background: Kidney disease (lupus nephritis [LN]) constitutes a feature of systemic lupus erythematosus (SLE) in up to 50 - 70% of patients with the disease. Although most LN patients are suitable for renal transplantation when they develop end stage renal disease (ESRD), the risk of recurrence of LN post-transplantation can be as high as 30%. Since the outcomes of renal transplantation in ESRD-LN patients has not been adequately studied in South Africa, the present study aims to retrospectively explore the aforementioned objective in a single centre. Methodology: The study was designed as a retrospective descriptive study of patients with LN transplanted in the renal unit of Groote Schuur Hospital, Cape Town from 1st January 2004 to 31st December 2013. Results: There were 454 patients who were transplanted in the study period of which 15/454 (3.3%) had LN. The M:F ratio of LN patients was 1:14, mean age was 25±10 years, all were known with class- IV LN and 10/15 (66.7%) received graft from a cadaveric donor. Immunosuppression was initiated in 7/15 (46.7%) with combination of cyclosporine and azathioprine; in 2/15 (13.3%) with tacrolimus and azathioprine and in 6/15 (40.0%) with Tacrolimus and MMF. All patients received corticosteroids. Recurrence of LN was seen in one patient (6.7%) who developed class V LN. Graft rejection was diagnosed in 10/15 cases (66.7%) with types of rejection noted to be acute cellular rejection in 6/15 (40%), antibody mediated rejection 1/15 (6.7%) and chronic rejection in 3/15 (20%). ESRD occurred in 3 patients (20%) with causes from antibody mediated rejection (6.7%), chronic allograft nephropathy (6.7%) and renal artery thrombosis (6.7%). Mean time to ESRD was 16.0 months. Five deaths (33.3%) occurred from sepsis in 3/15 (20%), pulmonary embolism; 1/15 (6.7%) and progressive ESRD after non-acceptance to the chronic dialysis program; 1/15 (6.7%). Mean time to death was 44.1 months. Conclusion: This study shows that recurrence of LN in the graft kidney is uncommon in South Africa. However, effort to reduce high rates of rejection and improve graft and patient survival still needs to be studied.

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Thesis (Ph.D.) in Biochemistry and Molecular Biology--University of Maine, 2007. / Includes vita. Includes bibliographical references (leaves 121-158).

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No description available.

Lu, Jianxin. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 218-241). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Wong, Tsz Yan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 140-159). / Abstracts in English and Chinese. / Acknowledgements --- p.I / Abbreviations --- p.III / Abstract --- p.V / 摘要 --- p.VIII / Publications --- p.XI / Table of Contents --- p.XII / Chapter Chapter 1: --- General Information / Chapter 1.1 --- Characteristics and Prevalence of SLE --- p.1 / Chapter 1.2 --- Diagnosis of SLE --- p.2 / Chapter 1.3 --- Assessment of Disease Activity --- p.4 / Chapter 1.4 --- Causes of SLE --- p.4 / Chapter 1.4.1 --- Genetic Factors --- p.4 / Chapter 1.4.2 --- Hormonal Factors --- p.6 / Chapter 1.4.3 --- Environment Factors --- p.7 / Chapter 1.5 --- Drug Treatments of SLE --- p.8 / Chapter 1.6 --- Immunological Dysregulation in SLE --- p.9 / Chapter 1.6.1 --- Types and Properties of Lymphocytes --- p.9 / Chapter 1.6.2 --- Cytokines and Chemokines --- p.14 / Chapter 1.6.3 --- Toll-like Receptors --- p.17 / Chapter 1.6.4 --- Intracellular Signal Transduction Pathways --- p.21 / Chapter 1.7 --- Objectives of Our study --- p.25 / Chapter Chapter 2: --- Material and Methods / Chapter 2.1 --- Materials --- p.27 / Chapter 2.1.1 --- SLE Patients and Control Subjects --- p.27 / Chapter 2.1.2 --- Reagents for cell culture --- p.28 / Chapter 2.1.3 --- Reagents for Flow Cytometry --- p.30 / Chapter 2.1.4 --- Reagents for Phosphorylation State Analysis --- p.33 / Chapter 2.1.5 --- Reagents for Total RNA Extraction --- p.35 / Chapter 2.1.6 --- Reagents for Polymerase Chain Reaction (PCR) --- p.36 / Chapter 2.1.7 --- Reagents for Gel Electrophoresis --- p.38 / Chapter 2.1.8 --- Reagents for Real-Time Polymerase Chain Reaction --- p.39 / Chapter 2.1.9 --- Other Reagents --- p.40 / Chapter 2.2 --- Methods --- p.41 / Chapter 2.2.1 --- Preparation of Plasma and Purification of Peripheral Blood Mononuclear Cells (PBMC) from EDTA-Blood --- p.41 / Chapter 2.2.2 --- Immunophenotyping of Cell Surface Molecules by Flow Cytometry --- p.42 / Chapter 2.2.3 --- Immunophenotyping of Intracellular Molecules by Flow Cytometry --- p.43 / Chapter 2.2.4 --- Phosphorylation State Analysis of Intracellular Signaling Molecules --- p.43 / Chapter 2.2.5 --- Cytometric Bead Array of Cytokines and Chemokines --- p.44 / Chapter 2.2.6 --- Total RNA Extraction from PBMC --- p.46 / Chapter 2.2.7 --- Reverse Transcription of the Extracted Total RNA --- p.46 / Chapter 2.2.8 --- Real-time Polymerase Chain Reaction --- p.46 / Chapter 2.2.9 --- Enzyme-Linked Immunosorbent Assay (ELISA) --- p.48 / Chapter 2.2.10 --- Enzyme-Linked Immunosorbent Spot (ELISPOT) --- p.48 / Chapter 2.2.11 --- Statistical Analysis --- p.49 / Chapter Chapter 3: --- B Cell Chemokine CXCL13 in SLE / Chapter 3.1 --- Introduction --- p.49 / Chapter 3.2 --- Methods --- p.52 / Chapter 3.3 --- Results --- p.52 / Chapter 3.3.1 --- Characteristics of SLE Patients and Control Subjects --- p.52 / Chapter 3.3.2 --- Expression of Plasma CXCL13 --- p.53 / Chapter 3.3.3 --- Gene Expression of TNF-α in PBMC --- p.53 / Chapter 3.3.4 --- Expression of sTNFRl in Plasma --- p.53 / Chapter 3.4 --- Discussion --- p.57 / Chapter Chapter 4: --- T lymphocyte Co-stimulatory Molecule CD26 in SLE / Chapter 4.1 --- Introduction --- p.61 / Chapter 4.2 --- Methods --- p.64 / Chapter 4.3 --- Results --- p.66 / Chapter 4.3.1 --- Characteristic of SLE Patients and Control Subjects --- p.66 / Chapter 4.3.2 --- Expression of Human CD26 in Plasma --- p.66 / Chapter 4.3.3 --- "Cell Surface Expression of CD26 on Monocytes, Th, Tc plus Ts, B and iNKT Lymphocytes" --- p.66 / Chapter 4.3.4 --- Circulating Number of iNKT Lymphocytes --- p.67 / Chapter 4.4 --- Discussion --- p.71 / Chapter Chapter 5: --- Thl7 Lymphocytes and Expression of IL-17 in SLE / Chapter 5.1 --- Introduction --- p.76 / Chapter 5.2 --- Methods --- p.78 / Chapter 5.3 --- Results --- p.79 / Chapter 5.3.1 --- Characteristics of SLE Patients and Control Subjects --- p.79 / Chapter 5.3.2 --- Ex vivo production of IL-17A from PBMC --- p.79 / Chapter 5.3.3 --- Circulating Number of Thl7 Lymphocytes --- p.79 / Chapter 5.4 --- Discussion --- p.82 / Chapter Chapter 6: --- Intracellular Mitogen Activated Protein Kinases in SLE / Chapter 6.1 --- Introduction --- p.87 / Chapter 6.2 --- Methods --- p.89 / Chapter 6.3 --- Results --- p.91 / Chapter 6.3.1 --- Characteristics of SLE Patients and Control Subjects --- p.91 / Chapter 6.3.2 --- Expression of Phospho-p38 MAPK in PBMC --- p.91 / Chapter 6.3.3 --- Expression of Phospho-ERK in PBMC --- p.92 / Chapter 6.3.4 --- Expression of Phospho-JNK in PBMC --- p.92 / Chapter 6.3.5 --- "Relative Percentage Increase of Phosphorylated p38 MAPK, ERK and JNK upon IL-18 Activation" --- p.93 / Chapter 6.4 --- Discussion --- p.104 / Chapter Chapter 7: --- Toll-like Receptors in SLE / Chapter 7.1 --- Introduction --- p.111 / Chapter 7.2 --- Methods --- p.113 / Chapter 7.3 --- Results --- p.115 / Chapter 7.3.1 --- Characteristics of SLE Patients and Control Subjects --- p.115 / Chapter 7.3.2 --- Expression of TLRl to TLR9 of PBMC --- p.115 / Chapter 7.3.3 --- Preliminary Results of Cytokine and Chemokine Expression Upon TLR Lignad Activation --- p.116 / Chapter 7.4 --- Discussion --- p.126 / Chapter Chapter 8: --- Conclusion and Future Perspectives --- p.133 / Appendix --- p.138 / References --- p.140