Noha Afifi

Glomerular expression and elevated serum Bcl-2 and Fas proteins in lupus nephritis : preliminary findings

Programmed cell death (apoptosis) is involved in glomerular injuries leading to glomerulonephritis. Bcl‐2 and Fas are proteins that promote cell survival and death, respectively. This study tests the hypothesis that lupus nephritis is associated with alterations of Bcl‐2 and Fas protein expression. Thirty‐six patients with lupus nephritis and 10 controls (normal individuals) were included in this study. Bcl‐2 and Fas positive cells were examined in kidney biopsies by immunohistochemistry. Bcl‐2 and Fas serum levels were evaluated by enzyme‐linked immunosorbent assay (ELISA). In the glomeruli of normal kidneys, Bcl‐2 and Fas proteins were completely absent. In lupus nephritis patients, glomerular expression of Bcl‐2 and Fas was seen in mesangial cells (1·3 ± 0·1 and 2·0 ± 0·1 for Bcl‐2 and Fas, respectively). Similarly, a statistically significantly higher Bcl‐2 (217·1 ± 85·9) and Fas (767·9 ± 271) serum levels were found in lupus patients compared to controls (148·6 ± 87, 550·3 ± 91 for Bcl‐2 and Fas, P < 0·05). A direct correlation between serum Bcl‐2 and Fas and chronicity index was also found. Compared to normal controls, lupus nephritis is associated with glomerular expression and elevated serum levels of Bcl‐2 and Fas proteins. These findings suggest possible roles for Bcl‐2 and Fas in glomerular injury during evolution of lupus nephritis. The diagnostic, prognostic and therapeutic ramifications of our findings are open to further investigation.

Dual effect of nitric oxide donor on adjuvant arthritis

The effect of medical use of NO donors on the pathogenesis of arthritis is still yet unclear. We investigated the effects of the NO donor, sodium nitroprusside (SNP), on the pathogenesis of adjuvant-induced arthritis in rats. Rats were given SNP intraperitoneally either from day 5 to day 14 (as a prophylactic protocol) or from day 16 to day 25 (as a therapeutic protocol) after inoculation of adjuvant. SNP administration, whether prophylactic or therapeutic, in doses of 0.1 and 1 mg/kg/d significantly aggravated pathogenesis of adjuvant arthritis in rats. SNP-treated rats showed significant (P<0.05) increase in arthritis index, hind paw volume, ankle joint diameter and hyperalgesia compared with control adjuvant arthritic rats. However, in adjuvant rats given the smallest dose of SNP (0.01 mg/kg/d), arthritis index, volume of hind paws, ankle joint diameter, body weight loss, and hyperalgesia were significantly lower than that of control adjuvant rats. After 30 d of the induction of adjuvant arthritis, TNF alpha levels exhibited insignificant changes either in control adjuvant rats or in rats given SNP compared with control non adjuvant rats. IL-10 levels in adjuvant control rats and adjuvant rats given 1 mg or 0.1 mg/kg/d from day 15 to day 25 were significantly lower than that of control non adjuvant rats. Histopathology examination of ankle joint showed that large doses of SNP (1 mg or 0.1 mg/kg/d) increased the mononuclear cells infiltration and erosion of cartilage induced by adjuvant while the infiltration of the inflammatory cells in the synovium of adjuvant rats treated with 0.01 mg/kg/d was minimal and the pannus was inhibited with alleviation of erosion of articular cartilage. Prophylactic small dose of SNP improved the histological status more than the therapeutic small dose. The present work reveals that SNP administration, either prophylactic or therapeutic, was deleterious in higher doses. However, the smallest dose used 0.01 mg/kg/d attenuates joint inflammation, hyperalgesia and body weight loss in adjuvant arthritic rats. These results suggest that small dose of NO donor may exert partial protective effects while the safety of the clinical use of NO donors, in higher doses, in patients with rheumatoid arthritis is questioned.

Enhancement of the anti‐inflammatory and anti‐arthritic effects of theophylline by a low dose of a nitric oxide donor or non‐specific nitric oxide synthase inhibitor

Background and purpose:  Although there are many new specific phosphodiesterase inhibitors with anti‐inflammatory activity, none have yet reached the market because of their low therapeutic efficacy. Our study was aimed to evaluate the anti‐inflammatory and anti‐arthritic effect of an established phosphodiesterase inhibitor, theophylline, and to investigate the effect of the nitric oxide (NO) donor, sodium nitroprusside (SNP) or NO synthase inhibitor, L‐NG‐monomethyl arginine (L‐NMMA) on its actions.

Regulatory T cell subsets in children with systemic lupus erythematosus

The aim of this work was to quantify CD4+CD25+Foxp3+ T cells (Tregs) in Egyptian children with SLE and to correlate these findings with their disease activity scores and drug therapy. We enrolled 37 Egyptian children with active SLE. Disease activity was assessed by measuring serum levels of anti-dsDNA antibody and by the SLEDAI scores. Twenty healthy children were also enrolled as normal controls. The CD4+CD25+, CD4+CD25bright, and CD4+CD25dim cells in patients were significantly increased in comparison to controls. There was no significant difference in the Foxp3 gated on CD4+CD25bright and CD4+CD25dim, but there was a significant increase when gated on CD4+CD25− and whole CD4+ cells in patients than controls. There was no significant difference among patients with different degrees of activity on different lines of treatments and their outcomes as regards all studied values. There was no significant correlation between SLEDAI score and any of the studied parameters except for a significant negative correlation with gated lymphocytes. There is increased expression of Foxp3 in CD4+ T cells mostly CD25− in Egyptian children with active SLE under corticosteroid treatment regardless of disease activity.

Influence of dipyridamole and its combination with NO donor or NO synthase inhibitor on adjuvant arthritis

The anti-arthritic and anti-inflammatory effects of dipyridamole and the possible involvement of NO in the dipyridamole action are not yet clear. The aim of this study was to evaluate the effects of dipyridamole alone and in combination with either the nitric oxide donor, sodium nitroprusside (SNP) or the non-selective nitric oxide synthase inhibitor, L-NG- monomethyl arginine (L-NMMA), on pathogenesis of adjuvant-induced arthritis model in rats. The results of the present work showed that prophylactic administration of dipyridamole alone and dipyridamole administration in combination with either low dose of SNP or L-NMMA significantly ameliorated pathogenesis of adjuvant arthritis in rats as evidenced by significant decrease in arthritis index, hind paws volume, loss of body weight, hyperalgesia compared with control vehicle (1% DMSO) treated adjuvant arthritic rats. Inflammatory cellular infiltrate in synovium of ankle joint and pannus formation were also markedly inhibited. Interleukin-10(IL-10) levels were significantly increased in these groups of animals. In contrast, a high dose of SNP counteracted the anti-inflammatory and anti-arthritic effects of dipyridamole. The inhibitory effect of therapeutic administration of dipyridamole alone on adjuvant arthritis syndrome was not significantly different from that of vehicle administration. In conclusion, dipyridamole has prophylactic but not therapeutic anti-arthritic and anti-inflammatory effects that appear to be dependent on inhibition of NO synthase. A synergistic combination between dipyridamole and NO synthase inhibitor or low dose of NO donor may have prophylactic and therapeutic values in autoimmune diseases like RA.

The efficacy of hepatitis B vaccination program in upper Egypt: Flow cytometry and the evaluation of long term immunogenicity.

Anti‐HBs levels wanes with time. Many studies discussed the B cell response to HBV vaccine. However, the data about memory T cell response are limited. To evaluate the efficacy of hepatitis B vaccine via evaluating anti‐HBs levels and HBsAg specific memory T‐lymphocytes through descriptive study. The study was conducted in a tertiary care setting. This study included 440 vaccinated persons during infancy. Group I: 6 to less than 10 years old; Group II: 10 to less than 14 years old; Group III: 14 to less than 17 years old; Group IV: 17 years old. The serum samples were screened for HBV markers. Cytokines secretion by HBsAg‐specific memory CD45RO+ CD4+ T cells was measured after in vitro culture using flow cytometry. The mean titer of anti‐HBs was higher in group I in comparison to others (P‐value = 0.000 for each). IFN‐γ and IL‐4 secreted by memory CD4+ T cells were positive in all with anti‐HBs >100 mIU/ml, while positive in 87% and 75% of participants with anti‐HBs <10 mIU/ml and positive in 73% and 32% of participants with absent anti‐HBs. The percentage of cells secreting IFN‐γ and those secreting IL‐4 were higher among participants with serum anti‐HBs >100 mIU/ml than those having <10 mIU/ml or absent (P < 0.001 for each). Anti‐HBs positivity decreased with time since childhood vaccination. Breakthrough infections are rare in vaccinated persons. Hepatitis‐B vaccine is efficient in controlling HBV infection. Flow cytometry is a useful tool to assess the long term persistence of T cell memory after childhood vaccination. J. Med. Virol. 88:1567–1575, 2016.

Role of T-Helper 9 Cells in Chronic Hepatitis C-Infected Patients

Hepatitis C virus is a hepatotropic virus that is transmitted parenterally. Viral infections are usually associated with modulations of the immune cells, leading to enhanced viral survival and spreading, and accordingly, life-threatening complications. Recently, it has been proposed that a new subset of T-helper, named T-helper 9, is involved in the pathogenesis of different immunopathological conditions, such as allergies, tumors, and viral infections. Some studies reported a protective role, and others described a pathogenic potential for the T-helper 9 cells. Here, we present evidence that T-helper 9 cells are dynamically increased with increasing the pathogenic strategy for hepatitis C virus (HCV). Furthermore, viral clearance is associated with a decrease in T-helper 9. The increase in T-helper 9 was paralleled with an increase in its receptor expression. Taken together, our data suggest that T-helper 9 cells play an important role in the pathogenesis of HCV, and is directly associated with HCV-related complications.

485 Regulatory T Cells Subsets in Children with Sle

In the case of SLE, Treg deficiencies have been described in mouse models of SLE. However, there are somehow conflicting data in the literature on whether Treg cells in human SLE are numerically and/or functionally impaired. We aimed to quantify CD4+CD25+Foxp3+ T cells in children with SLE and to correlate these findings with their disease activity scores and drug therapy. We enrolled 37 pediatric SLE patients and 20 healthy children. The disease activity was assessed by measuring serum levels of anti-dsDNA antibody and by using scores of SLEDAI. The CD4+CD25+, CD4+CD25bright and CD4+CD25dim cells in patients were significantly increased than controls. There was no significant difference in the FoxP3% gated on CD4+CD25bright, CD4+CD25dim and CD4+CD25-cells in patients and controls and between different grades of activity, different lines of treatments and patients outcomes as regards all studied values. There was no significant correlation between any of studied parameters with SLEDAI score except gated lymphocytes which have significant negative correlation. The increase of CD4+ CD25+ T cells in pediatric patients with active SLE may be a result of increased usage of corticosteroids that affect the phenotype of the T cells without affection on its regulatory suppression function indicated by FoxP3.